研究者総覧

井上 裕康 (イノウエ ヒロヤス)

  • 研究院生活環境科学系食物栄養学領域 教授
メールアドレス:
inouecc.nara-wu.ac.jp
Last Updated :2021/04/15

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学位

  • 医学博士, 大阪大学
  • 医科学修士, 大阪大学

研究キーワード

  • ミラクリン 味覚受容体 シクロオキシゲナーゼ プロスタグランジン 核内受容体 分子栄養学 生活習慣病 miraculin taste receptor cyclooxygenase prostaglandin nuclear receptor molecular nutrition life-style related diseases 

研究分野

  • ライフサイエンス, 医化学
  • ライフサイエンス, 食品科学
  • 人文・社会, 家政学、生活科学

経歴

  • 1994年 - 2004年 :国立循環器病センター研究所薬理部分子薬理研究室室長
  • 1994年 - 2004年 :Division Chief, Division of Molecular Pharmacololgy, Dept. of Pharmacology,
  • 2004年 -:奈良女子大学生活環境学部食物栄養学科教授
  • 2004年 -:Professor, Dept. of Food Science and Nutrition, Faculty of Human Life and Environment, Nara Women's University
  • 1987年 - 1994年 :国立循環器病センター研究所生化学部酵素化学研究室研究員
  • 1987年 - 1994年 :Investigator, Division of Enzyme Chemistry, Dept of Biochemistry,
  • 1991年 - 1993年 :ダラスサウスウエスタンメディカルセンター、ケンタッキー大学医学部生化学教室
  • 1991年 - 1993年 :Southwestern Medical Center at Dalla, Dept. of Biochemistry, University of Kentucky Medical Shool (Professor Louis B. Hersh)
  • 1986年 - 1987年 :日本学術振興会奨励研究員(ドクターコース)
  • (研究留学、Professor Louis B. Hersh)
  • National Cardiovascular Center Research Institute
  • National Cardiovascular Center Research Institute

学歴

  • - 1987年 大阪大学 医学研究科 生理系 日本国
  • - 1987年 大阪大学 Medical School (Doctor Course) Dept. Nutrition & Physiological chemistry

委員歴

  • 2012年04月 脂溶性ビタミン総合研究委員会 委員 脂溶性ビタミン総合研究委員会 society

    学協会

  • 2011年11月 日本ポリフェノール学会 理事 日本ポリフェノール学会 society

    学協会

  • 2009年11月 日本脂質生化学会 幹事 日本脂質生化学会 society

    学協会

  • 2006年05月 日本ビタミン学会 評議員 日本ビタミン学会 society

    学協会

  • 2005年 日本生化学会 評議員 日本生化学会 society

    学協会

  • 2005年 日本栄養食糧学会 評議員 日本栄養食糧学会 society

    学協会

  • 2010年05月 - 2012年04月 日本家政学会 関西支部庶務幹事 日本家政学会 society

    学協会

  • 2011年 日本味と匂い学会 評議員 日本味と匂い学会 society

    学協会

受賞

  • ICOFF2011 poster award, 2011年
  • ICOFF2011 poster award, 2011年
  • 実用化のための可能性試験(FS), 2006年, JPN
  • 森永奉仕会賞, 2005年, JPN
  • ネスレ栄養科学会議 助成金, 2005年, JPN
  • Research grant from the Nestle Nutrition Council, Japan, 2005年
  • 日本ビタミン学会奨励賞, 2002年, JPN
  • 日本血管作動物質学会学会賞, 1996年, JPN

論文

  • Lactobacillus helveticus-MIKI-020 enhances hepatic FGF21 expression and decreases the core body temperature during sleep in mice

    Kiriyama K; Goto T; Yamamoto H; Ara T; Takahashi H; Jheng HF; Nomura W; Inoue H; Nakata R; Kawada T

    2019年03月, Journal of Functional Foods, 54, 529 - 535

  • A phytol-enriched diet activates PPAR‐α in the liver and brown adipose tissue to ameliorate obesity-induced metabolic abnormalities.

    An JY; Jheng HF; Nagai H; Sanada K; Takahashi H; Iwase M; Watanabe N; Kim YI; Teraminami A; Takahashi N; Nakata R; Inoue H; Seno S; Matsuda H; Kawada T; Goto T

    Scope: Peroxisome proliferator-activated receptor alpha (PPAR-α) is a ligand-activated transcription factor that regulates lipid and carbohydrate metabolism. We investigate the effects of naturally occurring PPAR-α agonists, phytol, and its metabolite phytanic acid, on obesity-induced metabolic disorders using a mouse model. Methods and results: A luciferase reporter assay shows that phytanic acid potently activates PPAR-α among PPAR subtypes. In high-fat-diet-induced, severely obese mice, a phytol-enriched diet increases phytanic acid levels in the liver and adipose tissue, where PPAR-α is abundantly expressed. A phytol-enriched diet ameliorates severe obesity and the related metabolic abnormalities of white adipose tissue. Moreover, the expression of PPAR-α target genes in the liver and brown adipose tissue is enhanced by a phytol-enriched diet, suggesting that phytol and phytanic acid activate PPAR-α in these organs. We confirm that phytanic acid treatment induced PPAR-α target gene expression in both primary hepatocytes and brown adipocytes from wild-type mice, but not in these cells from PPAR-α-deficient mice. Conclusion: A phytol-enriched diet may increase phytanic acid levels in the liver and brown adipocytes, thereby activating PPAR-α in these organs and ameliorating obesity-induced metabolic diseases., 2018年03月01日, Mol. Nutr. Food Res., 62 (6), e1700688, doi

    研究論文(学術雑誌)

  • Erratum: Tumor-Associated Macrophage-Induced Invasion and Angiogenesis of Human Basal Cell Carcinoma Cells by Cyclooxygenase-2 Induction (Journal of Investigative Dermatology (2009) 129(4) (1016–1025)(S0022202X15342901)(10.1038/jid.2008.310))

    Jeng-Wei Tjiu; Jau-Shiuh Chen; Chia-Tung Shun; Sung-Jan Lin; Yi-Hua Liao; Chia-Yu Chu; Tsen-Fang Tsai; Hsien-Ching Chiu; Yang-Shia Dai; Hiroyasu Inoue; Pan-Chyr Yang; Min-Liang Kuo; Shiou-Hwa Jee

    Tumor-associated macrophages (TAMs) and cyclooxygenase-2 (COX-2) are associated with invasion, angiogenesis, and poor prognosis in many human cancers. However, the role of TAMs in human basal cell carcinoma (BCC) remains elusive. We found that the number of TAMs infiltrating the tumor is correlated with the depth of invasion, microvessel density, and COX-2 expression in human BCC cells. TAMs also aggregate near COX-2 expressing BCC tumor nests. We hypothesize that TAMs might activate COX-2 in BCC cells and subsequently increase their invasion and angiogenesis. TAMs are a kind of M2 macrophage derived from macrophages exposed to Th2 cytokines. M2-polarized macrophages derived from peripheral blood monocytes were cocultured with BCC cells without direct contact. Coculture with the M2 macrophages induced COX-2-dependent invasion and angiogenesis of BCC cells. Human THP-1 cell line cells, after treated with phorbol myristate acetate (PMA), differentiated to macrophages with M2 functional profiles. Coculture with PMA-treated THP-1 macrophages induced COX-2-dependent release of matrix metalloproteinase-9 and subsequent increased invasion of BCC cells. Macrophages also induced COX-2-dependent secretion of basic fibroblast growth factor and vascular endothelial growth factor-A, and increased angiogenesis in BCC cells. © 2009 The Society for Investigative Dermatology., 2018年02月01日, Journal of Investigative Dermatology, 129 (4), 1016 - 1025, doi;pubmed

    研究論文(学術雑誌)

  • レスベラトロール研究の進展

    中田理恵子; 井上裕康

    2018年, 生化学, 90 (6), 529 - 532

  • Rice Koji extract enhances lipid metabolism through prolifeator-activated receptor alpha (PPARα) activation in mouse liver.

    Takahashi H; Chi H-Y; Mohri S; Kamakari K; Nakata K; Ochijo N; Nakata R; Inoue H; Goto T; Kawada T

    Koji is made from grains fermented with Aspergillus oiyzae and is essential for the production of many traditional Japanese foods. Many previous studies have shown that koji contributes to the improvement of dyslipidemia. However, little is known regarding the underlying mechanism of this effect. Furthermore, the compound contributing to the activation of lipid metabolism is unknown. We demonstrated that rice koji extract (RKE) induces the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR alpha) target genes, which promotes lipid metabolism in murine hepatocytes. This effect was not observed in PPAR alpha-KO hepatocytes. We also demonstrated that RKE contained linolenic acid (LIA), oleic acid (OA), and hydroxyoctadecadienoic acids (HODEs), which activate PPAR alpha, using LC-MS analysis. Our findings suggest that RKE, containing LIA, OA, and HODEs, could be valuable in improving dyslipidemia via PPAR alpha activation., 2016年11月, J. Agric. Food Chem., 64 (46), 8848 - 8856, doi;web_of_science

    研究論文(学術雑誌)

  • Single ingestion of soy β-conglycinin induces increased postprandial circulating FGF21 levels exerting beneficial health effects.

    Hashizume T; Kato A; Tanaka T; Miyoshi S; Itoh N; Nakata R; Inoue H; Oikawa A; Nakai Y; Shimizu M; Inoue J; Sato R

    Soy protein beta-conglycinin has serum lipid-lowering and anti-obesity effects. We showed that singleingestion of beta-conglycinin after fasting alters gene expression in mouse liver. A sharp increase in fibroblast growth factor 21 (FGF21) gene expression, which is depressed by normal feeding, resulted in increased postprandial circulating FGF21 levels along with a significant decrease in adipose tissue weights. Most increases in gene expressions, including FGF21, were targets for the activating transcription factor 4 (ATF4), but not for peroxisome proliferator-activated receptor a. Overexpression of a dominant-negative form of ATF4 significantly reduced beta-conglycinin-induced increases in hepatic FGF21 gene expression. In FGF21-deficient mice, beta-conglycinin effects were partially abolished. Methionine supplementation to the diet or primary hepatocyte culture medium demonstrated its importance for activating liver or hepatocyte ATF4-FGF21 signaling. Thus, dietary beta-conglycinin intake can impact hepatic and systemic metabolism by increasing the postprandial circulating FGF21 levels., 2016年06月, Sci. Rep., 6, 28183, doi;web_of_science

    研究論文(学術雑誌)

  • 9-Oxo-10(E),12(Z),15(Z)-Octadecatrienoic Acid Activates Peroxisome Proliferator-Activated Receptor α in Hepatocytes.

    Takahashi H; Kamakari K; Goto T; Hara H; Mohri S; Suzuki H; Shibata D; Nakata R; Inoue H; Takahashi N; Kawada T

    2015年09月, Lipids, doi;pubmed

  • Metabolomics reveal 1-palmitoyl lysophosphatidylcholine production by peroxisome proliferator-activated receptor α.

    Takahashi H; Goto T; Yamazaki Y; Kamakari K; Hirata M; Suzuki H; Shibata D; Nakata R; Inoue H; Takahashi N; Kawada T

    2015年02月, Journal of lipid research, 56 (2), 254 - 265, doi;pubmed

  • The 4’-hydroxyl group of resveratrol is functionally important for direct activation of PPARα

    Yoshie Takizawa; Rieko Nakata; Kiyoshi Fukuhara; Hiroshi Yamashita; Hideo Kubodera; Hiroyasu Inoue

    Long-term moderate consumption of red wine is associated with a reduced risk of developing lifestyle-related diseases such as cardiovascular disease and cancer. Therefore, resveratrol, a constituent of grapes and various other plants, has attracted substantial interest. This study focused on one molecular target of resveratrol, the peroxisome proliferator activated receptor α (PPARα). Our previous study in mice showed that resveratrol-mediated protection of the brain against stroke requires activation of PPARα; however, the molecular mechanisms involved in this process remain unknown. Here, we evaluated the chemical basis of the resveratrol-mediated activation of PPARα by performing a docking mode simulation and examining the structure-activity relationships of various polyphenols. The results of experiments using the crystal structure of the PPARα ligand-binding domain and an analysis of the activation of PPARα by a resveratrol analog 4-phenylazophenol (4-PAP) in vivo indicate that the 4'-hydroxyl group of resveratrol is critical for the direct activation of PPARα. Activation of PPARα by 5 μM resveratrol was enhanced by rolipram, an inhibitor of phosphodiesterase (PDE) and forskolin, an activator of adenylate cyclase. We also found that resveratrol has a higher PDE inhibitory activity (IC50 = 19 μM) than resveratrol analogs trans-4-hydroxystilbene and 4-PAP (IC50 = 27-28 μM), both of which has only 4'-hydroxyl group, indicating that this 4'-hydroxyl group of resveratrol is not sufficient for the inhibition of PDE. This result is consistent with that 10 μM resveratrol has a higher agonistic activity of PPARα than these analogs, suggesting that there is a feedforward activation loop of PPARα by resveratrol, which may be involved in the long-term effects of resveratrol in vivo., 2015年, PLoS One, 10 (3), e0120865, 国際誌, doi;pubmed;pmc

  • Resveratrol Targets in Inflammation

    Hiroyasu Inoue; Rieko Nakata

    Resveratrol, a constituent of grapes and various other plants, has been an attractive compound for biomedical studies because moderate long-term drinking of red wine is associated with a reduced risk of lifestyle-related diseases, such as cardiovascular diseases and cancer. Resveratrol is as a phytoalexin, cyclooxygenase (COX) suppressor, and an activator of peroxisome proliferator-activated receptor (PPAR) and SIRT1. As a major phytoalexin, resveratrol is produced by plants in response to various environmental stresses, such as pathogens and ultraviolet (UV) radiation, and promotes resistance to these stresses. A similar active ingredient, salicylic acid (SA), is also produced by plants. Aspirin, acetylated SA, is a major nonsteroidal anti-inflammatory drug (NSAID) because it inhibits COX activity in humans. The jasmonic acid (JA) pathway in plants and the COX pathway in humans are both defense systems against environmental stresses and involve lipid mediators derived from phospholipids. We can hypothesize that there is a molecular basis for the mutually beneficial relationship between plants and humans, which is important for understanding the mode of action of resveratrol in inflammation. Here we provide a review of the studies on resveratrol, especially with respect to the role of COX and PPAR in inflammation., 2015年, ENDOCRINE METABOLIC & IMMUNE DISORDERS-DRUG TARGETS, 15 (3), 186 - 195, doi;web_of_science

    研究論文(学術雑誌)

  • The role of AMPK and PPARγ1 in exercise-induced lipoprotein lipase in skeletal muscle.

    井上 裕康; Sasaki T; Nakata R; Inoue H; Shimizu M; Inoue J; Sato R

    Exercise can effectively ameliorate type 2 diabetes and insulin resistance. Here we show that the mRNA levels of one of peroxisome proliferator-activated receptor (PPAR) family members, PPAR gamma 1, and genes related to energy metabolism, including PPAR gamma coactivator-1 protein-1 alpha (PGC-1 alpha) and lipoprotein lipase (LPL), increased in the gastrocnemius muscle of habitual exercise-trained mice. When mice were intraperitoneally administered an AMP-activated protein kinase (AMPK) activator 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR), the mRNA levels of the aforementioned three genes increased in gastrocnemius muscle. AICAR treatment to C2C12 differentiated myotubes also increased PPAR gamma 1 mRNA levels, but not PPAR alpha and -delta mRNA levels, concomitant with increased PGC-1 alpha mRNA levels. An AMPK inhibitor, compound C, blocked these AICAR effects. AICAR treatment increased the half-life of PPAR gamma 1 mRNA nearly threefold (4-12 h) by activating AMPK. When C2C12 myoblast cells infected with a PPAR gamma 1 expression lentivirus were differentiated into myotubes, PPAR gamma 1 overexpression dramatically increased LPL mRNA levels more than 40-fold. In contrast, when PPAR gamma 1 expression was suppressed in C2C12 myotubes, LPL mRNA levels were significantly reduced, and the effect of AICAR on increased LPL gene expression was almost completely blocked. These results indicated that PPAR gamma was intimately involved in LPL gene expression in skeletal muscle and the AMPK-PPAR gamma 1 pathway may play a role in exercise-induced LPL expression. Thus, we identified a novel critical role for PPAR gamma 1 in response to AMPK activation for controlling the expression of a subset of genes associated with metabolic regulation in skeletal muscle., 2014年05月, Am J Physiol Endocrinol Metab., 306 (9), E1085 - E1092, doi;web_of_science

    研究論文(学術雑誌)

  • Resveratrol and Cardiovascular Disease (invited review)

    井上 裕康; Rieko Nakata; Hiroyasu Inoue

    2014年, Current Nutrition Reports, Springer, 3, 163 - 169

  • Upregulation of eNOS、SIRT1 and autophagy-related genes by repeated treatment with resveratrol in human umbilical vein endothelial cells.

    井上 裕康; Takizawa Y; Kosuge Y; Awaji H; Tamura E; Takai A; Yanai T; Yamamoto R; Kokame K; Miyata T; Nakata R; Inoue H

    Resveratrol, a polyphenolic phytoalexin found in red wine and various plants, has been reported to up-regulate the expression of endothelial NO synthase (eNOS) in human umbilical vein endothelial cells (HUVEC). However, this effect was neither long term in nature nor physiologically relevant at the concentration of resveratrol studied. In the present study, we investigated the effects of repeated treatments with a lower concentration of resveratrol on the expression of genes in HUVEC. The expression levels of eNOS and silent mating type information regulation 2 homologue 1 (SIRT1) were up-regulated in HUVEC by repeated treatments with 1 μm-resveratrol for 6 d, but not with fenofibrate. Moreover, resveratrol treatment increased the expression of autophagy-regulated genes such as γ-aminobutyric acid A receptor-associated protein (GABARAP), microtubule-associated protein 1 light chain 3B (LC3B) and autophagy-related protein 3 (ATG3), the radical scavenger activity-related metallothionein-1X (MT1X) gene and the anti-inflammatory activity-related annexin A2 (ANXA) gene. In addition, resveratrol treatment down-regulated the expression of the cell-cycle checkpoint control RAD9 homologue B (RAD9B) gene. These results indicate the beneficial effects of resveratrol on the cardiovascular system. © The Authors 2013., 2013年12月28日, British J. Nutrition, 110 (12), 2150 - 2155, doi;pubmed

    研究論文(学術雑誌)

  • DHA attenuates postprandial hyperlipidemia via activating PPARα in intestinal epithelial cells.

    井上 裕康; Kimura R; Takahashi N; Lin S; Goto T; Murota K; Nakata R; Inoue H; Kawada T

    It is known that peroxisome proliferator-activated receptor (PPAR)alpha, whose activation reduces hyperlipidemia, is highly expressed in intestinal epithelial cells. Docosahexaenoic acid (DHA) could improve postprandial hyperlipidemia, however, its relationship with intestinal PPAR alpha activation is not revealed. In this study, we investigated whether DHA can affect postprandial hyperlipidemia by activating intestinal PPAR alpha using Caco-2 cells and C57BL/6 mice. The genes involved in fatty acid (FA) oxidation and oxygen consumption rate were increased, and the secretion of triacylglyceride (TG) and apolipoprotein B (apoB) was decreased in DHA-treated Caco-2 cells. Additionally, intestinal FA oxidation was induced, and TG and apoB secretion from intestinal epithelial cells was reduced, resulting in the attenuation of plasma TG and apoB levels after oral administration of olive oil in DHA-rich oil-fed mice compared with controls. However, no increase in genes involved in FA oxidation was observed in the liver. Furthermore, the effects of DHA on intestinal lipid secretion and postprandial hyperlipidemia were abolished in PPAR alpha knockout mice. In conclusion, the present work suggests that DHA can inhibit the secretion of TG from intestinal epithelial cells via PPAR alpha activation, which attenuates postprandial hyperlipidemia., 2013年12月, J Lipid Res., 54 (12), 3258 - 3268, doi;web_of_science

    研究論文(学術雑誌)

  • Secretion of miraculin through the function of a signal peptide conserved in the Kunitz-type soybean trypsin inhibitor family

    Ayako Takai; Makiko Satoh; Tomomi Matsuyama; Akane Ito; Rieko Nakata; Takashi Aoyama; Hiroyasu Inoue

    Miraculin, a glycoprotein that modifies sour tastes into sweet ones, belongs to the Kunitz-type soybean trypsin inhibitor (STI) family. To clarify the functional relation of miraculin with Kunitz-type STIs, we investigated its subcellular localization and trypsin inhibitory activity. In transgenic Arabidopsis thaliana, miraculin, fused to yellow fluorescent protein, localized to and outside the plasma membrane depending on the putative secretion signal peptide. When transgenic seedlings were cultured in liquid medium, miraculin was present in the supernatant only after cellulase treatment. No trypsin inhibitory activity was detected in native or recombinant miraculin. In conclusion, miraculin is secreted outside the plasma membrane through the function of a signal peptide, conserved in Kunitz-type STIs, whereas its trypsin inhibitory activity may be lost during its evolution. (C) 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., 2013年06月, FEBS LETTERS, 587 (12), 1767 - 1772, doi;web_of_science

    研究論文(学術雑誌)

  • Evaluation of food-derived functional ingredients according to activation of PPAR and suppression of COX-2 expression

    Rieko Nakata; Yoshie Takizawa; Ayako Takai; Hiroyasu Inoue

    The peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors belonging to the nuclear receptor family. They are considered molecular targets for the prevention of lifestyle-related diseases and are involved in the control of cyclooxygenase (COX)-2 expression. COX-2, the rate-limiting enzyme in prostaglandin biosynthesis, plays a key role in inflammation and circulatory homeostasis, and its expression is partly controlled by PPAR. We have identified several natural chemicals, such as resveratrol, that activate PPARs and suppress COX-2 expression. In this review, we provide an evaluation of food-derived functional ingredients that target PPARs and COX-2., 2013年, Food Science and Technology Research, 19 (3), 339 - 345, doi

  • レスベラトロール研究の現状

    井上 裕康; 中田 理恵子; 井上裕康

    2013年, 日本栄養士会雑誌, 56, 544 - 546

  • Recent Advances in the Study on Resveratrol

    Rieko Nakata; Satoru Takahashi; Hiroyasu Inoue

    Appropriate long-term drinking of red wine is associated with a reduced risk for lifestyle-related diseases such as cardiovascular disease and cancer, making resveratrol, a constituent of grapes and various other plants, an attractive compound to be studied. Historically, resveratrol has been identified as a phytoalexin, antioxidant, cyclooxygenase (COX) inhibitor, peroxisome proliferator-activated receptor (PPAR) activator, endothelial nitric oxide synthase (eNOS) inducer, silent mating type information regulation 2 homolog I (SIRT1) activator, and more. Despite scepticism concerning the biological availability of resveratrol, a growing body of in viva evidence indicates that resveratrol has protective effects in several stress and disease models. Here, we provide a review of the studies on resveratrol, especially with respect to COX, PPAR, and eNOS activities, and discuss its potential for promoting human health., 2012年03月, BIOLOGICAL & PHARMACEUTICAL BULLETIN, 35 (3), 273 - 279, doi;web_of_science

  • Farnesol, an isoprenoid, improves metabolic abnormalities in mice via both PPARα-dependent and -independent pathways.

    井上 裕康; Goto T; Kim YI; Funakoshi K; Teraminami A; Uemura T; Hirai S; Lee JY; Makishima M; Nakata R; Inoue H; Senju H; Matsunaga M; Horio F; Takahashi N; Kawada T

    Goto T, Kim YI, Funakoshi K, Teraminami A, Uemura T, Hirai S, Lee JY, Makishima M, Nakata R, Inoue H, Senju H, Matsunaga M, Horio F, Takahashi N, Kawada T. Farnesol, an isoprenoid, improves metabolic abnormalities in mice via both PPAR alpha-dependent and -independent pathways. Am J Physiol Endocrinol Metab 301: E1022-E1032, 2011. First published August 23, 2011; doi:10.1152/ajpendo.00061.2011.-Peroxisome proliferator-activated receptors (PPARs) control energy homeostasis. In this study, we showed that farnesol, a naturally occurring ligand of PPARs, could ameliorate metabolic diseases. Obese KK-Ay mice fed a high-fat diet (HFD) containing 0.5% farnesol showed significantly decreased serum glucose level, glucosuria incidence, and hepatic triglyceride contents. Farnesol-containing HFD upregulated the mRNA expressions of PPAR alpha target genes involved in fatty acid oxidation in the liver. On the other hand, farnesol was not effective in upregulating the mRNA expressions of PPAR gamma target genes in white adipose tissues. Experiments using PPAR alpha-deficient [(-/-)] mice revealed that the upregulation of fatty acid oxidation-related genes required PPAR alpha function, but the suppression of hepatic triglyceride accumulation was partially PPAR alpha dependent. In hepatocytes isolated from the wild-type and PPAR alpha (-/-) mice, farnesol suppressed triglyceride synthesis. In luciferase assay, farnesol activated both PPAR alpha and the farnesoid X receptor (FXR) at similar concentrations. Moreover, farnesol increased the mRNA expression level of a small heterodimer partner known as one of the FXR target genes and decreased those of sterol regulatory element-binding protein-1c and fatty acid synthase in both the wildtype and PPAR alpha (-/-) hepatocytes. These findings suggest that farnesol could improve metabolic abnormalities in mice via both PPAR alpha-dependent and -independent pathways and that the activation of FXR by farnesol might contribute partially to the PPAR alpha-independent hepatic triglyceride content-lowering effect. To our knowledge, this is the first study on the effect of the dual activators of PPAR alpha and FXR on obesity-induced metabolic disorders., 2011年11月, Am J Physiol Endocrinol Metab., 301 (5), E1022 - E1032, doi;web_of_science

    研究論文(学術雑誌)

  • A possible function of resveratrol and essential oil-derived chemicals for prevention of life-style related diseases targeted to COX-2 and PPAR

    Hiroyasu Inoue; Yoshie Takizawa; Ayako Takai; Chisako Takashiba; Satomi Koeji; Chiharu Iwasa; Rieko Nakata

    2011年08月, CHEMISTRY AND PHYSICS OF LIPIDS, 164, S29 - S29, doi;web_of_science

  • Citronellol and Geraniol, Components of Rose Oil, Activate PPAR α and γ and Suppress Cyclooxygenase-2 Expression

    井上 裕康; M. Katsukawa; R. Nakata; S. Koeji; K. Hori; S. Takahashi; H.Inoue

    We evaluated the effects of rose oil on the peroxisome proliferator-activated receptor (PPAR) and cyclooxygenase-2 (COX-2). Citronellol and geraniol, the major components of rose oil, activated PPAR alpha and gamma, and suppressed LPS-induced COX-2 expression in cell culture assays, although the PPAR gamma-dependent suppression of COX-2 promoter activity was evident only with citronellol, indicating that citronellol and geraniol were the active components of rose oil., 2011年05月, Biosci. Biotech. Biochem., 75 (5), 1010 - 1012, doi;web_of_science

    研究論文(学術雑誌)

  • Activation of peroxisome proliferator-activated receptor-alpha stimulates both differentiation and fatty acid oxidation in adipocytes

    Tsuyoshi Goto; Joo-Young Lee; Aki Teraminami; Yong-Il Kim; Shizuka Hirai; Taku Uemura; Hiroyasu Inoue; Nobuyuki Takahashi; Teruo Kawada

    Peroxisome proliferator-activated receptor-alpha (PPAR alpha) is a dietary lipid sensor, whose activation results in hypolipidemic effects. In this study, we investigated whether PPAR alpha activation affects energy metabolism in white adipose tissue (WAT). Activation of PPAR alpha by its agonist (beza-fibrate) markedly reduced adiposity in KK mice fed a high-fat diet. In 3T3-L1 adipocytes, addition of GW7647, a highly specific PPAR alpha agonist, during adipocyte differentiation enhanced glycerol-3-phosphate dehydrogenase activity, insulin-stimulated glucose uptake, and adipogenic gene expression. However, triglyceride accumulation was not increased by PPAR alpha activation. PPAR alpha activation induced expression of target genes involved in FA oxidation and stimulated FA oxidation. In WAT of KK mice treated with bezafibrate, both adipogenic and FA oxidation-related genes were significantly upregulated. These changes in mRNA expression were not observed in PPAR alpha-deficient mice. Bezafibrate treatment enhanced FA oxidation in isolated adipocytes, suppressing adipocyte hypertrophy. Chromatin immunoprecipitation (ChIP) assay revealed that PPAR alpha was recruited to promoter regions of both adipogenic and FA oxidation-related genes in the presence of GW7647 in 3T3-L1 adipocytes.(jlr) These findings indicate that the activation of PPAR alpha affects energy metabolism in adipocytes, and PPAR alpha activation in WAT may contribute to the clinical effects of fibrate drugs.-Goto, T., J-Y. Lee, A. Teraminami, Y-I. Kim, S. Hirai, T. Uemura, H. Inoue, N. Takahashi, and T. Kawada. Activation of peroxisome proliferator-activated receptor-alpha stimulates both differentiation and fatty acid oxidation in adipocytes. J. Lipid Res. 2011. 52: 873-884., 2011年05月, JOURNAL OF LIPID RESEARCH, 52 (5), 873 - 884, doi;web_of_science

    研究論文(学術雑誌)

  • Gene and protein analysis of brain derived neurotrophic factor expression in relation to neurological recovery induced by an enriched environment in a rat stroke model

    Kenji Hirata; Yuji Kuge; Chiaki Yokota; Akina Harada; Koichi Kokame; Hiroyasu Inoue; Hidekazu Kawashima; Hiroko Hanzawa; Yuji Shono; Hideo Saji; Kazuo Minematsu; Nagara Tamaki

    Although an enriched environment enhances functional recovery after ischemic stroke, the mechanism underlying this effect remains unclear. We previously reported that brain derived neurotrophic factor (BDNF) gene expression decreased in rats housed in an enriched environment for 4 weeks compared to those housed in a standard cage for the same period. To further clarify the relationship between the decrease in BDNF and functional recovery, we investigated the effects of differential 2-week housing conditions on the mRNA of BDNF and protein levels of proBDNF and mature BDNF (matBDNF). After transient occlusion of the right middle cerebral artery of male Sprague-Dawley rats, we divided the rats into two groups: (1) an enriched group housed multiply in large cages equipped with toys, and (2) a standard group housed alone in small cages without toys. Behavioral tests before and after 2-week differential housing showed better neurological recovery in the enriched group than in the standard group. Synaptophysin immunostaining demonstrated that the density of synapses in the pen-infarct area was increased in the enriched group compared to the standard group, while infarct volumes were not significantly different. Real-time reverse transcription polymerase chain reaction. Western blotting and immunostaining all revealed no significant difference between the groups. The present results suggest that functional recovery cannot be ascribed to an increase in matBDNF or a decrease in proBDNF but rather to other underlying mechanisms. (C) 2011 Elsevier Ireland Ltd. All rights reserved., 2011年05月, NEUROSCIENCE LETTERS, 495 (3), 210 - 215, doi;web_of_science

    研究論文(学術雑誌)

  • Gene expression associated with an enriched environment after transient focal ischemia

    Yuji Shono; Chiaki Yokota; Yuji Kuge; Shinsuke Kido; Akina Harada; Koichi Kokame; Hiroyasu Inoue; Mariko Hotta; Kenji Hirata; Hideo Saji; Nagara Tamaki; Kazuo Minematsu

    Recent studies have demonstrated that animals housed in an enriched environment after an experimental stroke obtained a better functional outcome than those housed in a standard cage; however, little is known about the gene expression associated with this functional recovery. The purpose of the present study was to elucidate the expression of genes in an enriched environment after experimental stroke in the ischemic and non-ischemic sides of the cortices. Transient focal brain ischemia was produced by the occlusion of the right middle cerebral artery (t-MCAO) in male Sprague Dawley rats. The rats were divided into 3 groups: ischemic rats housed in the enriched environment, ischemic rats housed in standard cages, and non-ischemic rats in standard cages. Four weeks after t-MCAO, the rats were sacrificed and gene expression was examined. Motor function was improved in ischemic rats housed in the enriched environment compared with those in standard cages; however, there were no significant differences in the size of the infarct area between the ischemic rats in the enriched environment and those in standard cages. Decreases in the expression of Egr-1, -2, and BDNF mRNA in both sides of the cortices were detected in rats housed in the enriched environment, indicating that gene expression was altered throughout the brain at 4 weeks after transient focal ischemia. (C) 2010 Elsevier B.V. All rights reserved., 2011年02月, BRAIN RESEARCH, 1376, 60 - 65, doi;web_of_science

    研究論文(学術雑誌)

  • Citral, a component of lemongrass oil, activates PPARα and γ and suppresses COX-2 expression

    井上 裕康; Katsukawa M; Nakata R; Takizawa T; Hori, K; Takahashi, S; Inoue, H

    Lemongrass is a widely used herb as a food flavoring, as a perfume, and for its analgesic and anti-inflammatory purposes; however, the molecular mechanisms of these effects have not been elucidated. Previously, we identified carvacrol from the essential oil of thyme as a suppressor of cyclooxygenase (COX)-2, a key enzyme for prostaglandin synthesis, and also an activator of peroxisome proliferator-activated receptor (PPAR), a molecular target for "lifestyle-related" diseases. In this study, we evaluated the essential oil of lemongrass using our established assays for COX-2 and PPARs. We found that COX-2 promoter activity was suppressed by lemongrass oil in cell-based transfection assays, and we identified citral as a major component in the suppression of COX-2 expression and as an activator of PPAR alpha and gamma. PPAR gamma-dependent suppression of COX-2 promoter activity was observed in response to citral treatment In human macrophage-like U937 cells, citral suppressed both U'S-induced COX-2 mRNA and protein expression, dose-dependently. Moreover, citral induced the mRNA expression of the PPAR alpha-responsive carnitine palmitoyltransferase 1 gene and the PPAR gamma-responsive fatty acid binding protein 4 gene, suggesting that citral activates PPAR alpha and gamma, and regulates COX-2 expression. These results are important for understanding the anti-inflammatory and anti-lifestyle-related disease properties of lemongrass. (C) 2010 Elsevier B.V. All rights reserved., 2010年11月, BBA - Molecular and Cell Biology of Lipids, 1801 (11), 1214 - 1220, doi;web_of_science

    研究論文(学術雑誌)

  • SIRT1 modulates expression of matrix metalloproteinases in human dermal fibroblasts

    K. Ohguchi; T. Itoh; Y. Akao; H. Inoue; Y. Nozawa; M. Ito

    P>Background SIRT1, an NAD+-dependent histone/protein deacetylase, controls a broad range of cellular functions. Objectives We examined if SIRT1 is involved in the regulation of matrix metalloproteinase (MMP) expression in human dermal fibroblasts. Methods We studied the effect of inhibition of SIRT1 by specific inhibitor and small interfering RNA (siRNA) on MMP-1 and MMP-3 expression in human dermal fibroblasts. Results Treatment with a potent and selective inhibitor of SIRT1, EX-527, increased the basal expression levels of MMP-1 and MMP-3 proteins. Knockdown of endogenous SIRT1 by siRNA led to increased expression of MMP-1 and MMP-3 at both mRNA and protein levels. SIRT1 knockdown also upregulated MMP protein induction caused by an inflammatory cytokine, interleukin (IL)-1 beta. Moreover, treatment with a SIRT1 activator, resveratrol, significantly suppressed IL-1 beta-mediated induction of MMP-1, which was attenuated by pretreatment with EX-527. Finally, MMP-1 promoter activity was increased by EX-527 in cells treated with or without IL-1 beta. Conclusions Our findings suggest that SIRT1 exerts a negative regulatory role in the production of MMP-1 and MMP-3 in human dermal fibroblasts., 2010年10月, BRITISH JOURNAL OF DERMATOLOGY, 163 (4), 689 - 694, doi;web_of_science

    研究論文(学術雑誌)

  • Ethanolic extracts of Brazilian red propolis promote adipocyte differentiation through PPAR gamma activation

    Akio Iio; Kenji Ohguchi; Hiroyasu Inoue; Hiroe Maruyama; Yoko Araki; Yoshinori Nozawa; Masafumi Ito

    Aim of the study: The aim of present study was to investigate the effects of ethanolic extracts of red propolis (EERP) on adipogenesis and evaluate the molecular basis for their anti-obesity effects. Materials and methods: We tested whether EERP alone could induce differentiation of 313-L1 cells, regulate the expression of adipocyte-specific genes and reverse inhibitory effects of TNF-alpha on their differentiation. Next, we performed a luciferase reporter gene assay to test whether EERP could enhance transcriptional activities of PPAR gamma and adiponectin promoter activities. Results: EERP strongly induced differentiation of 313-L1 preadipocytes into adipocytes, and enhanced the PPAR gamma transcriptional activity and adiponectin promoter activity. In addition, EERP attenuated the inhibitory effect of TNF-alpha on adipocyte differentiation and adiponectin production in mature adipocytes. Conclusion: The present study indicates that EERP enhance differentiation of 31341 adipocytes in part by its potency of PPAR gamma activation and are capable of reversing inhibitory effects of TNF-alpha on adipocyte differentiation and adiponectin expression. These results suggest the value of EERP as a diet supplement for prevention and treatment of obesity and obesity-associated disorders. (C) 2010 Elsevier GmbH. All rights reserved., 2010年10月, PHYTOMEDICINE, 17 (12), 974 - 979, doi;web_of_science

    研究論文(学術雑誌)

  • Vaticanol C, a resveratrol tetramer, activates PPARα and PPARβ/δ in vitro and in vivo

    井上 裕康; Tsukamoto T; Nakata R; Tamura E; Kosuge Y; Kariya A; Katsukawa M; Mishima S; Ito, T; Iinuma M; Akao Y; NozawaY; Arai Y; Namura S; Inoue H

    Background: Appropriate long-term drinking of red wine is associated with a reduced risk of cardiovascular disease. Resveratrol, a well-known SIRT1 activator is considered to be one of the beneficial components contained in red wine, and also developed as a drug candidate. We previously demonstrated that resveratrol protects brain against ischemic stroke in mice through a PPAR alpha-dependent mechanism. Here we report the different effects of the oligomers of resveratrol. Methods: We evaluated the activation of PPARs by epsilon-viniferin, a resveratrol dimer, and vaticanol C, a resveratrol tetramer, in cell-based reporter assays using bovine arterial endothelial cells, as well as the activation of SIRT1. Moreover, we tested the metabolic action by administering vaticanol C with the high fat diet to wild-type and PPAR alpha-knockout male mice for eight weeks. Results: We show that vaticanol C activates PPAR alpha and PPAR beta/delta in cell-based reporter assays, but does not activate SIRT1. epsilon-Viniferin shows a similar radical scavenging activity as resveratrol, but neither effects on PPARs and SIRT-1. Eight-week intake of vaticanol C with a high fat diet upregulates hepatic expression of PPAR alpha-responsive genes such as cyp4a10, cyp4a14 and FABP1, and skeletal muscle expression of PPAR beta/delta-responsive genes, such as UCP3 and PDK4 ( pyruvate dehydrogenase kinase, isoform 4), in wild-type, but not PPAR alpha-knockout mice. Conclusion: Vaticanol C, a resveratrol tetramer, activated PPAR alpha and PPAR beta/delta in vitro and in vivo. These findings indicate that activation of PPAR alpha and PPAR beta/delta by vaticanol C may be a novel mechanism, affording beneficial effects against lifestyle-related diseases., 2010年05月, Nutrition & Metabolism, 7 (46), doi;web_of_science

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  • Carvacrol, a component of thyme oil, activates PPARα and γ, and suppresses COX-2 expression

    井上 裕康; Hotta M; Nakata R; Katsukawa M; Hori K; Takahashi S; Inoue H

    Cyclooxygenase-2 (COX-2), the rate-limiting enzyme in prostaglandin biosynthesis, plays a key role in inflammation and circulatory homeostasis. Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors belonging to the nuclear receptor superfamily and are involved in the control of COX-2 expression, and vice versa. Here, we show that COX-2 promoter activity was suppressed by essential oils derived from thyme, clove, rose, eucalyptus, fennel, and bergamot in cell-based transfection assays using bovine arterial endothelial cells. Moreover, from thyme oil, we identified carvacrol as a major component of the suppressor of COX-2 expression and an activator of PPAR alpha and gamma. PPAR gamma-dependent suppression of COX-2 promoter activity was observed in response to carvacrol treatment. In human macrophage-like U937 cells, carvacrol suppressed lipopolysaccharide-induced COX-2 mRNA and protein expression, suggesting that carvacrol regulates COX-2 expression through its agonistic effect on PPAR alpha. These results may be important in understanding the antiinflammatory and antilifestyle-related disease properties of carvacrol.-Hotta, M., R. Nakata, M. Katsukawa, K. Hori, S. Takahashi, and H. Inoue. Carvacrol, a component of thyme oil, activates PPAR alpha and gamma and suppresses COX-2 expression. J. Lipid Res. 2010. 51: 132-139., 2010年01月, J. Lipid Res., 51 (1), 132 - 139, doi;web_of_science

    研究論文(学術雑誌)

  • 特殊なPPAR活性化能を示す香辛料シナモンバーク精油

    井上 裕康; 勝川 路子; 中田 理恵子; 滝澤 祥恵; 井上 裕康

    2010年, 日本味と匂学会誌, 17 (3), 203 - 206

  • Vascular endothelial growth factor-C (VEGF-C) promotes angiogenesis by induction of COX-2 in leukemic cells via the VEGF-R3/JNK/AP-1 pathway

    Ming-Hsien Chien; Chia-Chi Ku; Gunnar Johansson; Min-Wei Chen; Michael Hsiao; Jen-Liang Su; Hiroyasu Inoue; Kuo-Tai Hua; Lin-Hung Wei; Min-Liang Kuo

    Vascular endothelial growth factor (VEGF)-C is recognized as a tumor lymphangiogenic factor based on the effects of activated VEGF-R3 on lymphatic endothelial cells. Many tumor cells express VEGF-R3 but the function of this receptor in tumor cells is largely unknown. It has been reported that the VEGF-C/VEGF-R3 axis is activated in subsets of leukemia patients. Herein, we have shown that VEGF-C induces angiogenic activity in the tube formation assay invitro and Matrigel plug assay in vivo by upregulating an angiogenic factor, cyclooxygenase-2 (COX-2), through VEGF-R3 in the human acute myeloid leukemia (AML) cell line, THP-1. COX-2 induction by VEGF-C was also observed in other VEGF-R3(+) human AML cell lines (U937 and HL60). Moreover, immunohistochemical analysis of bone marrow specimens of 37 patients diagnosed with AML revealed that VEGF-C expression in specimens was associated with the expression of COX-2 (P < 0.001). The manner by which signaling pathways transduced by VEGF-C is responsible for COX-2 upregulation was further investigated. Blocking the p42/44 mitogen-activated protein kinase (MAPK) pathway with the MAPK kinase inhibitor, PD 98059, failed to inhibit VEGF-C-mediated COX-2 expression. However, VEGF-C-induced COX-2 upregulation was effectively abolished by overexpression of dominant-negative c-Jun N-terminal kinase (JNK) or treatment with the JNK inhibitor, SP 600125. VEGF-C induced JNK-dependent nuclear translocation of c-Jun. Furthermore, chromatin immunoprecipitation and reporter assays revealed that VEGF-C enhanced c-Jun binding to the cyclic adenosine 3',5'-monophosphate-response element of the COX-2 promoter and induced COX-2 expression. In sum, the data herein highlight the pathogenic role of VEGF-C in leukemia via regulation of angiogenesis through upregulation of COX-2., 2009年12月, CARCINOGENESIS, 30 (12), 2005 - 2013, doi;web_of_science

    研究論文(学術雑誌)

  • Tumor-Associated Macrophage-Induced Invasion and Angiogenesis of Human Basal Cell Carcinoma Cells by Cyclooxygenase-2 Induction

    Jeng-Wei Tjiu; Jau-Shiuh Chen; Chia-Tung Shun; Sung-Jan Lin; Yi-Hua Liao; Chia-Yu Chu; Tsen-Fang Tsai; Hsien-Ching Chiu; Yang-Shia Dai; Hiroyasu Inoue; Pan-Chyr Yang; Min-Liang Kuo; Shiou-Hwa Jee

    Tumor-associated macrophages (TAMs) and cyclooxygenase-2 (COX-2) are associated with invasion, angiogenesis, and poor prognosis in many human cancers. However, the role of TAMs in human basal cell carcinoma (BCC) remains elusive. We found that the number of TAMs infiltrating the tumor is correlated with the depth of invasion, microvessel density, and COX-2 expression in human BCC cells. TAMs also aggregate near COX-2 expressing BCC tumor nests. We hypothesize that TAMs might activate COX-2 in BCC cells and subsequently increase their invasion and angiogenesis. TAMs are a kind of M2 macrophage derived from macrophages exposed to Th2 cytokines. M2-polarized macrophages derived from peripheral blood monocytes were cocultured with BCC cells without direct contact. Coculture with the M2 macrophages induced COX-2-dependent invasion and angiogenesis of BCC cells. Human THP-1 cell line cells, after treated with phorbol myristate acetate (PMA), differentiated to macrophages with M2 functional profiles. Coculture with PMA-treated THP-1 macrophages induced COX-2-dependent release of matrix metalloproteinase-9 and subsequent increased invasion of BCC cells. Macrophages also induced COX-2-dependent secretion of basic fibroblast growth factor and vascular endothelial growth factor-A, and increased angiogenesis in BCC cells., 2009年04月, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 129 (4), 1016 - 1025, doi;web_of_science

    研究論文(学術雑誌)

  • Functional Expression of Miraculin, a Taste-Modifying Protein in Escherichia Coli

    Tomomi Matsuyama; Makiko Satoh; Rieko Nakata; Takashi Aoyama; Hiroyasu Inoue

    Miraculin isolated from red berries of Richadella dulcifica, a native shrub of West Africa, has the unusual property of modifying a sour taste into a sweet one. This homodimer protein consists of two glycosylated polypeptides that are cross-linked by a disulfide bond. Recently, functional expression of miraculin was reported in host cells with the ability to glycosylate proteins, such as lettuce, tomato and the microbe Aspergillus oryzae, but not Escherichia coli. Thus, a question remains as to whether glycosylation of miraculin is essential for its taste-modifying properties. Here we show that recombinant miraculin expressed in E. coli has taste-modifying properties as a homodimer, not as a monomer, indicating that glycosylation is not essential for the taste-modifying property., 2009年04月, JOURNAL OF BIOCHEMISTRY, 145 (4), 445 - 450, doi;web_of_science

    研究論文(学術雑誌)

  • Functional Expression of Miraculin, a Taste-Modifying Protein in Arabidopsis thaliana and Escherichia coli

    Makiko Satoh; Tomomi Matsuyama; Rieko Nakata; Takashi Aoyama; Hiroyasu Inoue

    2009年02月, CHEMICAL SENSES, 34 (2), J14 - J14, doi;web_of_science

  • シロイヌナズナにおけるミラクリン-YFP融合タンパク質の発現系構築

    井上 裕康; 佐藤 麻紀子; 中田 理恵子; 青山卓史; 井上裕康

    2009年, 日本味と匂学会誌, 16 (3), 293 - 294

  • COX-2発現抑制とPPAR活性化を指標にした香辛料成分の機能性評価

    井上 裕康; 勝川 路子; 中田 理恵子; 井上 裕康

    2009年, 日本味と匂学会誌, 16 (3), 683 - 686

  • p38 MAPK mediates COX-2 gene expression by corticosterone in cardiomyocytes

    Haipeng Sun; Beibei Xu; Hiroyasu Inoue; Qin M. Chen

    Recent work from our laboratory found that corticosteroids induce transcriptional activation of cyclooxygenase-2 (COX-2) gene in cardiomyocytes. Here we report that COX-2 gene promoter mutation studies indicate a role of cAMP response element-binding protein (CREB) in corticosterone-induced COX-2 gene expression. Corticosterone causes activation of p38 MAPK and subsequent CREB phosphorylation at serine 133 in cardiomyocytes. The inhibitors of p38 MAPK, SB202190 and S13203580, block corticosterone from inducing CREB phosphorylation and COX-2 gene expression while dominant-negative p38 MAPK or CREB prevents corticosterone from activating COX-2 promoter. Corticosterone does not induce p38 MAPK activation or COX2 expression in cardiac fibroblasts or HEK293 cells transfected with glucocorticoid receptor, suggesting that p38 MAPK activation is cell specific and necessary for corticosterone-induced COX-2 expression in cardiomyocytes. While glucocorticoid receptor antagonist mifepristone inhibits COX-2 gene induction by corticosterone, mifepristone fails to inhibit p38 MAPK activation or CREB phosphorylation. In contrast, inhibition of p38 MAPK does not prevent corticosterone from activating glucocorticoid receptor. Our data suggest that two parallel signaling pathways, glucocorticoid receptor and p38 MAPK, act in concert to regulate the expression of COX-2 gene in cardiomyocytes. (c) 2008 Published by Elsevier Inc., 2008年11月, CELLULAR SIGNALLING, 20 (11), 1952 - 1959, doi;web_of_science

    研究論文(学術雑誌)

  • Corticosteroids induced COX-2 expression in cardiomyocytes: Role of glucocorticoid receptor and C/EBPβ

    井上 裕康; Sun H; Sheveleva E; Xu B; Inoue H; Bowden TG; Chen QM

    Psychological stress increases the level of glucocorticoids in the circulating system. We found that dexamethasone administration in adult mice elevates the expression of COX-2 in the myocardium. With isolated neonatal cardiomyocytes, corticosterone (CT) at physiologically relevant doses (0.01-1 mu M) induces the expression of COX-2 gene. The induction first appeared at 4 h and remained for at least 24 h with 1 mu M CT treatment. This response is likely cardiomyocyte cell type specific since CT did not induce COX-2 expression in cardiac fibroblasts and glucocorticoids are known to suppress the expression of COX-2 in lymphocytes and several organs. Corticosteroids, but not estrogen or progesterone, induce COX-2 expression. The glucocorticoid receptor (GR) antagonist mifepristone (MF) prevented CT from inducing COX-2 gene, suggesting a GR-dependent induction in cardiomyocytes. COX-2 gene promoter deletion and mutation studies indicate a role of CCAAT/enhancer binding protein-beta (C/EBP-beta) in CT-induced COX-2 gene expression. Chromatin immunoprecipitation assays revealed that CT caused the binding of both GR and C/EBP-beta to COX-2 promoter, while MF pretreatment blocked such binding. Coimmunoprecipitation experiments demonstrated that CT treatment induced the interaction of GR with C/EBP-beta. Small interfering RNA against C/EBP-beta prevented CT from activating COX-2 promoter or elevating COX-2 protein. Our data suggest that the interaction between GR and C/EBP-beta contributes to elevated COX-2 gene transcription by CT in cardiomyocytes., 2008年10月, Am J Physiol Cell Physiol., 295 (4), C915 - C922, doi;web_of_science

    研究論文(学術雑誌)

  • 誘導型シクロオキシゲナーゼ発現抑制を指標とした植物油の機能性評価

    井上 裕康; 堀田真理子; 中田理恵子

    2008年, 日本家政学会誌, 59, 373 - 378, doi

  • Transcriptional regulation of the COX-2 expression by nitric oxide in colon cancer cell lines

    Qiang Liu; Hiroyasu Inoue; Ratha Mahendran

    To determine the effect of nitric oxide (NO) on the cyclooxygenase-2 (COX-2) regulation in colon cancer cell lines we used the physiological NO donor GSNO. The proximal 6.6 kb of the COX-2 promoter was cloned into the pGL3 basic vector and the sequential deletion of the 6.6 kb COX-2 promoter generated promoter constructs of 4, 2.6, 1.9 and 0.9 kb. These constructs clearly show that the main regulatory region lies within 0.9 kb of the transcription start site. Therefore, constructs of the main transcription binding sites within this region namely CRE, NF-IL6 and NF-kappa B and mutations of these sites were used to monitor the transcriptional activation of COX-2. This study was performed on the colon cancer cell lines HCA7 and HCT 116 which have a differential expression of COX-2. There was no evidence that the luciferase activity is negatively affected by NO as was previously reported. The CRE and NF-IL6 binding sites within this region were responsible for the constitutive and physiological NO-induced COX-2 transcriptional activity in the HCA7 and HCT 116 cells. While NF-kappa B involvement was only observed in the HCT116 cells, the cell lines displayed increased NF-kappa B transcriptional activity after exposure to NO., 2008年01月, ONCOLOGY REPORTS, 19 (1), 269 - 274, web_of_science

    研究論文(学術雑誌)

  • シロイヌナズナを用いた組換えミラクリンの発現と大腸菌を用いた組換え体との比較評価

    井上 裕康; 佐藤麻紀子; 松山友美; 中田理恵子; 青山卓司; 井上裕康

    2008年, 日本味と匂学会誌, 15, 517 - 520

  • Regulation of Cox-2 by cyclic AMP response element binding protein in prostate cancer: Potential role for nexrutine

    Rita Ghosh; Gretchen E. Garcia; Katherine Crosbyy; Hiroyasu Inoue; Ian M. Thompson; Dean A. Troyer; Addanki P. Kumar

    We recently showed that Nexrutine(R), a Phellodendron amurense bark extract, suppresses proliferation of prostate cancer cell lines and tumor development in the transgenic adenocarcinoma of mouse prostate (TRAMP) model. Our data also indicate that the anti-proliferative effects of Nexrutine(R) are mediated in part by Akt and Cyclic AMP response element binding protein (CREB). Cyclooxygenase (Cox-2), a pro-inflammatory mediator, is a CREB target that induces prostaglandin E-2 (PGE(2)) and suppresses apoptosis. Treatment of LNCaP cells with Nexrutine(R) reduced tumor necrosis factor A-induced enzymatic as well as promoter activities of Cox-2. Nexrutine(R) also reduced the expression and promoter activity of Cox-2 in PC-3 cells that express high constitutive levels of Cox-2. Deletion analysis coupled with mutational analysis of the Cox-2 promoter identified CRE as being sufficient for mediating Nexrutine R response. Immunohistochemical analysis of human prostate tumors show increased expression of CREB and DNA binding activity in high-grade tumors (three-fold higher in human prostate tumors compared to normal prostate; P =.01). We have identified CREB-mediated activation of Cox-2 as a potential signaling pathway in prostate cancer which can be blocked with a nontoxic, cost-effective dietary supplement like Nexrutine(R), demonstrating a prospective for development of Nexrutine(R) for prostate cancer management., 2007年11月, NEOPLASIA, 9 (11), 893 - 899, doi;web_of_science

    研究論文(学術雑誌)

  • Helicobacter pylori VacA enhances prostaglandin E-2 production through induction of cyclooxygenase 2 expression via a p38 mitogen-activated protein Kinase/Activating transcription factor 2 cascade in AZ-521 cells

    Junzo Hisatsune; Eiki Yamasaki; Masaaki Nakayama; Daisuke Shirasaka; Hisao Kurazono; Yohtaro Katagata; Hiroyasu Inoue; Jiahuai Han; Jan Sap; Kinnosuke Yahiro; Joel Moss; Toshiya Hirayama

    Treatment of AZ-521 cells with Helicobacter pylori VacA increased cyclooxygenase 2 (COX-2) mRNA in a time- and dose-dependent manner. A p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, blocked elevation of COX-2 mRNA levels, whereas PD98059, which blocks the Erk1/2 cascade, partially suppressed the increase. Consistent with involvement of p38 MAPK,, VacA-induced accumulation of COX-2 mRNA was reduced in AZ-521 cells overexpressing a dominant-negative p38 MAPK (DN-p38). Phosphatidylinositol-specific phospholipase C, which inhibits VacA-induced p38 MAPK activation, blocked VacA-induced COX-2 expression. In parallel with COX-2 expression, VacA increased prostaglandin E-2 (PGE(2)) production, which was inhibited by SB203580 and NS-398, a COX-2 inhibitor. VacA-induced PGE2 Production was markedly attenuated in AZ-521 cells stably expressing DN-p38. VacA increased transcription of a COX-2 promoter reporter gene and activated a COX-2 promoter containing mutated NF-kappa B or NF-interieukin-6 sites but not a mutated cis-acting replication element (CRE) site, suggesting direct involvement of the activating transcription factor 2 (ATF-2)/CREB-binding region in VacA-induced COX-2 promoter activation. The reduction of ATF-2 expression in AZ-521 cells transformed with ATF-2-small interfering RNA duplexes resulted in suppression of COX-2 expression. Thus, VacA enhances PGE2 production by AZ-521 cells through induction of COX-2 expression via the p38 MAPK/ATF-2 cascade, leading to activation of the CRE site in the COX-2 promoter., 2007年09月, INFECTION AND IMMUNITY, 75 (9), 4472 - 4481, doi;web_of_science

    研究論文(学術雑誌)

  • Role of cyclooxygenase-2 in tetrahydrobiopterin-induced dopamine oxidation

    Sung-Wook Chae; Yeo Jin Bang; Kyeong-Man Kim; Kwang Youl Lee; Bok Yun Kan; Eun Mee Kim; Hiroyasu Inoue; Onyou Hwang; Hyun Jin Choi

    Dopamine is considered one of the main contributing factors in the induction of oxidative stress and selective dopaminergic neurodegeneration in Parkinson's disease. We have previously reported that tetrahydrobiopterin (BH4) leads to dopamine oxidation and renders dopamine-producing cells vulnerable. In the present study, we found that BH4 selectively upregulates cyclooxygenase-2 (COX-2) expression in dopaminergic cells. BH4 caused an induction of COX-2 mRNA, and a critical regulatory motif for BH4-induced transcriptional activation of COX-2 is CRE/AP-1. COX-2 can oxidize dopamine and cause oxidative stress, which is evidenced by the findings that significant increase in dopamine-chrome formation and protein carbonyl contents by BH4-induced COX-2 up-regulation, and the increases are abolished by COX-2 selective inhibitor meloxicam. Increased COX-2 promotes dopaminergic neurodegeneration in both SH-SY5Y cells and rat mesencephalic neurons. These data suggest that BH4-induced COX-2 expression is responsible for dopamine oxidation, leading to the preferential vulnerability of dopaminergic cells in Parkinson's disease. (c) 2007 Elsevier Inc. All rights reserved., 2007年08月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 359 (3), 735 - 741, doi;web_of_science

    研究論文(学術雑誌)

  • Hypertonic sodium choloride and mannitol induces COX-2 via different signaling pathways in mouse cortical collecting duct M-1 cells

    WonChung Lim; JinSup Jung; Youngkon Surh; Hiroyasu Inoue; YoungJoo Lee

    The kidney cortical collecting duct is an important site for the maintenance of sodium balance. Previous studies have shown that, in renal medullary cells, hypertonic stress induces expression of cyclooxygenase-2 (COX-2) via NF-kappa B activation, but little is known about COX-2 expression in response to hypertonicity in the cortical collecting duct. Therefore, we examined the mechanism of hypertonic induction of COX-2 in M-1 cells derived from mouse cortical collecting duct. Induction of COX-2 protein was detected within 6 h of treatment with hypertonic sodium chloride. The treatment also increased COX-2 mRNA accumulation in a cycloheximide-independent manner, suggesting that ongoing protein synthesis is not required for COX-2 induction. Using reporter plasmids containing 0.2-, 0.3-, and 1.5-kb fragments of the COX-2 promoter, we found that hypertonic induction of COX-2 was due to an increase in promoter activity. The COX-2-inductive effect of hypertonicity was inhibited by SB203580, indicating that the effect is mediated by p38 MAPK. Since p38 MAPK can activate NF-kappa B, we made point mutations in the NF-kappa B binding site within the COX-2 promoter. The mutations did not block the induction of COX-2 promoter activity by hypertonic sodium chloride, and hypertonic sodium chloride failed to activate NF-kappa B binding site-driven reporter gene constructs. In contrast, hypertonic mannitol activated NF-kappa B, indicating that hypertonic mannitol and hypertonic sodium chloride activate COX-2 by different mechanisms. Thus, induction of COX-2 expression in M-1 cells by hypertonic sodium chloride does not involve activation of NF-kappa B. Furthermore, the signal transduction pathways that respond to hypertonic stress vary for different osmolytes in cortical collecting duct cells. (c) 2007 Elsevier Inc. All rights reserved., 2007年05月, LIFE SCIENCES, 80 (22), 2085 - 2092, doi;web_of_science

    研究論文(学術雑誌)

  • Targeting the NF-kappa B pathway through pharmacological inhibition of IKK2 prevents human cytomegalovirus replication and virus-induced inflammatory response in infected endothelial cells

    Patrizia Caposio; Tiziana Musso; Anna Luganini; Hiroyasu Inoue; Marisa Gariglio; Santo Landolfo; Giorgio Gribaudo

    Endothelial cells are important reservoirs for human cytomegalovirus (HCMV) replication, dissemination and persistence. HCMV infection of endothelial cells has been associated with a proinflammatory response characterized by an increased expression of chemokines and adhesion molecules and modulation of angiogenesis. Many of the host proinflammatory genes augmented in HCMV-infected endothelial cells are regulated, at least in part, by the NF-kappa B pathway. HCMV is a potent activator of NF-kappa B through the IKK-I kappa B signaling axis. To explore whether inhibition of HCMV-induced NF-kappa B activation may interfere with the onset of virus-associated inflammatory response, we measured the effects of the specific IKK2 inhibitor AS602868 on the expression of a panel of proinflammatory genes in HUVEC cells infected with a clinical isolate. Treatment of infected HUVEC with AS602868 was shown to impair HCMV-induced NF-kappa B activity, IE gene expression, viral replication and to prevent HCMV-induced upregulation of ICAM-1, IL-8, RANTES, IP-10, I-TAC and COX-2 gene expression. Consistent with these results, HCMV-mediated upregulation of another NF-kappa B-dependent gene, the plasminogen inhibitor type-1, a regulatory factor of endothelial proliferation and angiogenesis, was abrogated by AS602868. These results suggest that inhibition of HCMV-induced IKK-NF-kappa B activation may be of interest to limit the virus-induced inflammatory response of infected endothelial cells. (c) 2006 Elsevier B.V. All rights reserved., 2007年03月, ANTIVIRAL RESEARCH, 73 (3), 175 - 184, doi;web_of_science

    研究論文(学術雑誌)

  • Regulation of hepatic cholesterol synthesis by a novel protein (SPF) that accelerates cholesterol biosynthesis

    Norihito Shibata; Kou-ichi Jishage; Makoto Arita; Miho Watanabe; Yosuke Kawase; Kiyotaka Nishikawa; Yasuhiro Natori; Hiroyasu Inoue; Hitoshi Shimano; Nobuhiro Yamada; Masafumi Tsujimoto; Hiroyuki Arai

    Supernatant protein factor (SPF) is a novel cholesterol biosynthesis- accelerating protein expressed in liver and small intestine. Here, we report on the physiological role of SPF by using Spf-deficient mice. Although plasma cholesterol levels were similar in chow-fed Spf(-/-) and wild-type (WT) mice, fasting significantly decreased plasma cholesterol levels in Spf(-/-) mice but not in WT mice. While fasting reduced hepatic cholesterol synthesis rate in WT mice, a more pronounced reduction was observed in Spf(-/-) mice. The expression of cholesterogenic enzymes was dramatically suppressed by fasting both in WT and Spf(-/-) mice. In contrast, hepatic SPF expression of WT mice was up-regulated by fasting in peroxisome proliferator-activated receptor alpha (PPAR-alpha)-dependent manner. These results indicate that in WT mice, the decrease of hepatic cholesterol synthesis under fasting conditions is at least in part compensated by SPF up-regulation. Fibrates, which function as a PPAR-alpha agonist and are widely used as hypotriglycemic drugs, reduced hepatic cholesterol synthesis and plasma cholesterol levels by approximately one-half in Spf(-/-) mice but not in WT mice. These findings suggest that co-administration of fibrates and an SPF inhibitor may reduce not only plasma triglyceride but also cholesterol levels, indicating that SPF is a promising hypocholesterolemic drug target., 2006年12月, FASEB JOURNAL, 20 (14), 2642 - +, doi;web_of_science

    研究論文(学術雑誌)

  • Refolding and activation of human prorenin expressed in Escherichia coli: Application of recombinant human renin for inhibitor screening

    Saori Takahashi; Hironobu Ogasawara; Takayuki Watanabe; Masanori Kumagai; Hiroyasu Inoue; Kazuyuki Hori

    Human prorenin was expressed in Escherichia coli as a fusion protein of thioredoxin. The chimeric protein, which accumulated insoluble inclusion bodies, was solubilized in 4M guanidine-HCI and refolded by an arginine-detergent buffer system and by systematic dialysis. The refolded fusion prorenin was activated by trypsin. The antiserum against human kidney renin specifically inhibited the recombinant human renin activity. Using the recombinant human renin, we screened its inhibitory activity in fermented soybean paste (miso) and demonstrated that miso contained renin inhibitory activity derived from soybean. The IC50 values for soybean and steamed soybean extracts were determined to be 1.9 and 1.6 mg/ml, respectively. This is the first demonstration of renin inhibitory activity in miso and soybean., 2006年12月, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70 (12), 2913 - 2918, doi;web_of_science

    研究論文(学術雑誌)

  • Cox-2 is regulated by Toll-like receptor-4 (TLR4) signaling: Role in proliferation and apoptosis in the intestine

    Masayuki Fukata; Anli Chen; Arielle Klepper; Suneeta Krishnareddy; Arunan S. Vamadevan; Lisa S. Thomas; Ruliang Xu; Hiroyasu Inoue; Moshe Arditi; Andrew J. Dannenberg; Maria T. Abreu

    Background & Aims: We recently showed that mice deficient in Toll-like receptor 4 (TLR4) or its adapter molecule MyD88 have increased signs of colitis compared with wild-type (WT) mice after dextran sodium sulfate (DSS)-induced injury. We wished to test the hypothesis that cyclooxygenase 2 (Cox2)-derived prostaglandin E-2 (PGE(2)) is important in TLR4-related mucosal repair. Methods: Cox-2 expression was analyzed by real-time polymerase chain reaction, immunohistochemistry, Western blotting, and luciferase reporter constructs. Small interfering RNA was used to inhibit expression of MyD88. TLR4-/- or WT mice were given 2.5% DSS for 7 days. Proliferation and apoptosis were assessed using bromodeoxyuridine staining and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assays, respectively. PGE(2) was given orally to DSS-treated mice. Results: intestinal epithelial cell lines up-regulated Cox-2 expression in a TLR4- and MyD88-dependent fashion. Lipopolysaccharide-mediated stimulation of PGE(2) production was blocked by a selective Cox-2 inhibitor or small interfering RNA against MyD88. After DSS injury, Cox-2 expression increased only in WT mice. TLR4-/- mice have significantly reduced proliferation and increased apoptosis after DSS injury compared with WT mice. PGE(2) supplementation of TLR4-/- mice resulted in improvement in clinical signs of colitis and restoration of proliferation and apoptosis to WT values. The mechanism for improved epithelial repair may be through PGE(2)-dependant activation of the epidermal growth factor receptor. Conclusions: We describe an important link between TLR4 signaling and Cox-2 expression in the gut. TLR4 and MyD88 signaling are required for optimal proliferation and protection against apoptosis in the injured intestine. Although TLR4 signaling is beneficial in the short term, chronic signaling through TLR4 may lower the threshold for colitis-associated cancer., 2006年09月, GASTROENTEROLOGY, 131 (3), 862 - 877, doi;web_of_science

    研究論文(学術雑誌)

  • Oxidized-HDL3 modulates the expression of Cox-2 in human endothelial cells

    E Callegari; GD Norata; H Inoue; AL Catapano

    Modified high density lipoprotein (HDL) has been suggested to modulate endothelial expression of proinflammatory genes. Since oxidised HDL (Ox-HDL) has been found in atheromatous plaques and receptors for modified HDL are present on endothelial cells, we investigated the effect of Ox-HDL3 on the expression of Cox-1 or Cox-2. Ox-HDL3, increased Cox-2 mRNA and protein expression in endothelial cells while no effect on Cox-1 expression was observed. The intracellular pathways involved in this effect were investigated. The incubation with specific inhibitors of intracellular kinases showed that PI3K is mainly involved in the Ox-HDL3-dependent Cox-2 induction. Transient transfection experiments suggested that the NF-IL6 response element in the proximal promoter (-327 to 59) is involved in Ox-HDL3-mediated Cox-2 expression. These data suggest that Ox-HDL induce Cox-2 expression in endothelial cells through a PI3K/NF-IL6-dependent pathway., 2006年07月, INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, 18 (1), 209 - 213, web_of_science

    研究論文(学術雑誌)

  • Inhibitory effects of ginsenoside-Rb1 on activation of the 12-O-tetradecanoylphorbol 13-acetate-induced cyclooxygenase-2 promoter

    W Park; W Lim; J Cho; H Inoue; MR Rhyu; Y Lee

    We studied the inhibitory effects of ginsenoside-Rb1 (1) on 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced transcriptional activation of the cyclooxygenase-2 (COX-2) promoter. The suppressive activity of ginsenoside-Rbl was characterized using COX-2 promoter-driven luciferase reporter plasmids in a transient transfection system. Ginsenoside-Rb1 at 100 mu M inhibited TPA-induced transcriptional activation of the COX-2 promoter. To identify the cis-acting elements responsible for this inhibition, the effects of site-specific mutations in the COX-2 promoter region were examined. Inhibition by ginsenoside-Rb1 was not affected by mutations in nuclear factor-kappa B- or cAMP-responsive elements. However, the effects were abolished when the nuclear factor-interleukin-6 binding site was mutated, indicating that ginsenoside-Rbl exerts its effects via this element. In conclusion, ginsenoside-Rb1 inhibits TPA-induced COX-2 promoter activity through the nuclear factor interleukin-6 binding site and not through the nuclear factor-kappa B or cAMP-responsive elements., 2006年02月, PLANTA MEDICA, 72 (3), 272 - 275, doi;web_of_science

    研究論文(学術雑誌)

  • ET-18-O-CH3-induced apoptosis is causally linked to COX-2 upregulation in H-ras transformed human breast epithelial cells

    HK Na; H Inoue; YJ Surh

    Abnormally elevated expression of cyclooxygenase-2 (COX-2) has been frequently observed in transformed or malignant cells, and certain non-steroidal anti-inflammatory drugs with COX-2 inhibitory activity exert anti-neoplastic or chemopreventive effects. Contrary to this notion, we have found that a novel alkylphospholipid type antitumor agent ET-18-O-CH3 (1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine) induces COX-2 expression in H-ras transformed human breast epithelial cells (MCF10A-ras) while it causes apoptosis at the same concentration range. The addition of a selective COX-2 inhibitor SC-58635 and COX-2 gene knock down with the siRNA blocked ET-18-O-CH3-induced apoptosis, suggesting that COX-2 induction by this drug is causally linked to its apoptosis inducing activity. ET-18-O-CH3 enhanced the transcriptional activities of cyclic AMP response element which is a key regulator of COX-2 expression. 15-Deoxy-Delta-(12,14) prostaglandin J(2) is, an endogenous ligand for peroxisome proliferator-activated receptor gamma (PPAR gamma), has been known to possess proapoptotic potential in diverse cell types. ET-18-O-CH3 treatment resulted in elevated release of 15d-PGJ(2) and DNA binding and transcriptional activity of PPAR gamma. Based on these findings, it is likely that ET-18-O-CH3 induces COX-2 expression and production of 15d-PGJ2 which may mediate the ET-18-O-CH3-induced apoptosis in MCF10A-ras cells. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., 2005年11月, FEBS LETTERS, 579 (27), 6279 - 6287, doi;web_of_science

    研究論文(学術雑誌)

  • Heparanase is involved in angiogenesis in esophageal cancer through induction of cyclooxygenase-2

    T Okawa; Y Naomoto; T Nobuhisa; M Takaoka; T Motoki; Y Shirakawa; T Yamatsuji; H Inoue; M Ouchida; M Gunduz; M Nakajima; N Tanaka

    Purpose: Both heparanase and cyclooxygenase-2 (COX-2) are thought to play critical roles for tumor malignancy, including angiogenesis, although it is unknown about their relationship with each other in cancer progression. We hypothesized that they may link to each other on tumor angiogenesis. Experimental Design: The expressions of heparanase and COX-2 in 77 primary human esophageal cancer tissues were assessed by immunohistochemistry to do statistical analysis for the correlation between their clinicopathologic features, microvessel density, and survival of those clinical cases. Human esophageal cancer cells were transduced with heparanase c DNA and used for reverse transcription-PCR and Western blot to determine the expression of heparanase and COX-2. COX-2 promoter vector and its deletion/mutation constructs were also used along with transduction of heparanase c DNA for luciferase assay. Results: Heparanase and COX-2 protein expression exhibited a similar pattern in esophageal tumor tissues, and their expression correlated with tumor malignancy and poor survival. Their expression also revealed a significant correlation with high intratumoral microvessel density. Up-regulation of COX-2 mRNA and protein was observed in esophageal cancer cells transfected with heparanase cDNA. COX-2 promoter was activated after heparanase cDNA was transduced and the deletion/mutation of three transcription factor (cyclic AMP response element, nuclear factor-KB, and nuclear factor-interleukin-6) binding elements in COX-2 promoter strongly suppressed its activity. Conclusion: Our results suggest that heparanase may play a novel role for COX-2-mediated tumor angiogenesis., 2005年11月, CLINICAL CANCER RESEARCH, 11 (22), 7995 - 8005, doi;web_of_science

    研究論文(学術雑誌)

  • Nitric oxide upregulates the cyclooxygenase-2 expression through the cAMP-response element in its promoter in several cancer cell lines

    SW Park; MW Sung; DS Heo; H Inoue; SH Shim; KH Kim

    We previously showed that nitric oxide (NO) induces overexpression of cyclooxygenase-2 (COX-2) and production of prostaglandin E-2 in cancer cells. Here, we investigated the mechanisms by which NO induces COX-2 expression in cancer cells. We found that the cAMP-response element (CRE) is a critical factor in NO-induced COX-2 expression in all cells tested. We found that in cancer cells, three transcription factors (TFs) cAMP response element-binding protein (CREB), activating transcription factor-2 (ATF-2) and c-jun, bound the CRE in the COX-2 promoter, and their activities were increased by addition of the NO donor, S-nitroso-N-acetyl-D, L-penicillamine (SNAP). NO-induced activation of soluble guanylate cyclase (sGC), p38 and c-Jun NH2 terminal kinase (JNK) upregulated the three TFs, leading to COX-2 overexpression. Addition of dibutyryl-cGMP (db-cGMP) induced COX-2 expression in a manner similar to SNAP; this induction was blocked by a p38 inhibitor (SB202190), but not by a JNK inhibitor (SP600125). NO-induced cGMP was found to activate CREB and ATF-2 in a p38, but not c-jun- dependent manner,while NO induced JNK in a cGMP-independent manner, leading to subsequent activation of c-jun and ATF-2. These results suggest that the low concentrations of endogenous NO present in cancer cell may induce the expression of many genes, including COX-2, which promotes the growth and survival of tumor cells., 2005年10月, ONCOGENE, 24 (44), 6689 - 6698, doi;web_of_science

    研究論文(学術雑誌)

  • Hypertonic sodium chloride induction of cyclooxygenase-2 occurs independently of NF-kappa B and is inhibited by the glucocorticoid receptor in A549 cells

    WC Lim; M Park; JJ Bahn; H Inoue; YJ Lee

    Cellular response to a hypertonic environment is important for fluid clearance in the lung. Hypertonicity modulates prostaglandin synthesis by influencing cyclooxygenase-2 (COX-2) expression in tissues such as liver and kidney via a mitogen-activated protein kinase (MAPK)-dependent pathway. However, little is known about COX-2 expression in response to hypertonicity in the lung. COX-2 mRNA accumulation induced by hypertonic NaCl was detected after I h of treatment, and COX-2 mRNA continued to accumulate until 18 h, the longest time point examined, in human alveolar epithelial A549 cells. This induction was a transcriptional event that occurred in the absence of the protein synthesis inhibitor cycloheximide and was the result of enhanced promoter activity, as examined with the use of full-length COX-2 promoter-driven reporter plasmids. The induction of COX-2 expression by hypertonic NaCl did not require the activation of NF-kappa B. The p38 MAPK inhibitor, SB203580, or MEK1/2 inhibitor, U0126, inhibited hypertonic induction of COX-2 expression. We examined whether the hypertonic induction of COX-2 was under the influence of glucocorticoid; we found that COX-2 promoter activity and mRNA and protein levels were depressed by dexamethasone and antagonized by the glucocorticoid receptor (GR) antagonist RU486. Our data demonstrate that the induction of COX-2 expression by hypertonic NaCl occurs independently of NF-kappa B and is inhibited by the GR in A549 cells. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., 2005年10月, FEBS LETTERS, 579 (24), 5430 - 5436, doi;web_of_science

    研究論文(学術雑誌)

  • T-oligo treatment decreases constitutive and UVB-induced COX-2 levels through p53- and NF kappa B-dependent repression of the COX-2 promoter

    Marwaha, V; YH Chen; E Helms; S Arad; H Inoue; E Bord; R Kishore; RD Sarkissian; BA Gilchrest; DA Goukassian

    Chronically irradiated murine skin and UV light-induced squamous cell carcinomas overexpress the inducible isoform of cyclooxygenase (COX-2), and COX-2 inhibition reduces photocarcinogenesis in mice. We have reported previously that DNA oligonucleotides substantially homologous to the telomere 3'-overhang (T-oligos) induce DNA repair capacity and multiple other cancer prevention responses, in part through up-regulation and activation of p53. To determine whether T-oligos affect COX-2 expression, human newborn keratinocytes and fibroblasts were pretreated with T-oligos or diluent alone for 24 h, UV-irradiated, and processed for Western blotting. In both cell types, T-oligos transcriptionally down-regulated base-line and UV light-induced COX2 expression, coincident with p53 activation. In fibroblasts with wild type versus dominant negative p53 (p53(WT) versus p53(DN)), T-oligos decreased constitutive expression of a COX-2 reporter plasmid by>50%. Wethen examined NF kappa B, a known positive regulator of COX-2 transcription. In p53WT but not in p53DN fibroblasts and in human keratinocytes, T-oligos decreased readout of an NF kappa B promoter driven reporter plasmid and decreased NF kappa B binding to DNA. After T-oligo treatment and subsequent UV irradiation, binding of the transcriptional co-activator protein p300 to NF kappa B was decreased, whereas binding of p300 to p53 was increased. Human skin explants provided with T-oligos had markedly decreased COX-2 immunostaining both at base-line and post-UV light, coincident with increased p53 immunostaining. We conclude that T-oligos transcriptionally down-regulate COX-2 expression in human skin via activation and up-regulation of p53, at least in part by inhibiting NF kappa B transcriptional activation. Decreased COX-2 expression may contribute to the observed ability of T-oligos to reduce photocarcinogenesis., 2005年09月, JOURNAL OF BIOLOGICAL CHEMISTRY, 280 (37), 32379 - 32388, doi;web_of_science

  • Possible connection between "French Paradox" and PPAR

    H Inoue; H Takeuchi; T Matsubara; S Namura

    2005年09月, CHEMISTRY AND PHYSICS OF LIPIDS, 136 (2), 99 - 100, web_of_science

  • Oncogenic potential of MEK1 in rat intestinal epithelial cells is mediated via cyclooxygenase-2

    K Komatsu; FG Buchanan; S Katkuri; JD Morrow; H Inoue; M Otaka; S Watanabe; RN DuBois

    Background & Aims: The mitogen-activated protein kinase/extracellular signal-regulated protein kinase kinase (MEK) pathway plays an important role in the regulation of cell growth and differentiation. Constitutively active components of the MEK signaling cascade can induce oncogenic transformation in many cell systems. Downstream MEK signaling also plays an important role in the regulation of cyclooxygenase-2 (COX-2), which is known to be involved in colorectal cancer. Therefore, we determined the role of COX-2 on the oncogenic potential of MEK1 in nontransformed rat intestinal epithelial cells. Methods: Constitutively active MEK1 (CA-MEK) mutant transfected rat intestinal epithelial cells were established and tested for their ability to grow in soft agar and form tumors in vivo. The effect of CA-MEK on sodium butyrate (NaB)-induced apoptosis was evaluated by the Annexin V assay. The transcriptional activity and posttranscriptional stability of the COX-2 gene was determined by transient transfection with COX-2 reporter variants and by Northern analysis. To address the role of COX-2 in tumor growth in vivo, xenografted mice were treated with celecoxib (100 mg/kg) or vehicle. Results: CA-MEK transfected RIE-1 and IEC-6 cells formed colonies in soft agar and tumors in nude mice. These cells showed resistance to NaB-induced apoptosis and cell cycle arrest. MEK activation led to increased expression of COX-2, BCI-X-L, Mcl-1, and phosphorylated Bad and decreased expression of Bak. Along with elevated COX-2 levels, PGI(2) and PGE(2) levels were also increased. Pharmacologic inhibition of COX-2 inhibited MEK-induced tumor growth in vivo through enhanced apoptosis. Conclusions: COX-2 and its bioactive lipid products may play an important role in MEK-induced transformation., 2005年08月, GASTROENTEROLOGY, 129 (2), 577 - 590, doi;web_of_science

    研究論文(学術雑誌)

  • Bilateral induction of the S-100A9 gene in response to spreading depression is modulated by the cyclooxygenase-2 activity

    C Yokota; Y Kuge; H Inoue; N Tamaki; K Minematsu

    Cyclooxygenase-2 (COX-2) was reported to be induced in the infarcted human brain. Spreading depression (SD) is thought to play a role in this induction. In this study, we correlated the expression of SD-associated genes with COX-2 production in brains after SD. Rats were divided into 3 groups: rats that did not undergo SD (group I saline controls, n=7), rats that underwent unilateral SD as a result of KCl application (group II, n=9), and rats that were pretreated with the selective COX-2 inhibitor, JTE-522 3 h before the induction of SD (group III, n=7). The expression of the SD-associated genes, S-100A9, and mitogen-activated proteinkinase phosphatase (cpg21) was analyzed 2 h later using a cDNA array. In group II, COX-2 and cpg21 mRNA expression, as determined by RT-PCR, were significantly upregulated in the hemisphere undergoing SD. While the expression of S-100A9 mRNA was bilaterally upregulated in these animals, this expression was significantly reduced in group III, and was accompanied by reduced bilateral production of PGE(2). Thus, the bilateral induction of expression of the S-100A9 gene in response to SD was associated with COX-2 activation. (c) 2005 Elsevier B.V. All rights reserved., 2005年07月, JOURNAL OF THE NEUROLOGICAL SCIENCES, 234 (1-2), 11 - 16, doi;web_of_science

    研究論文(学術雑誌)

  • Helicobacter pylori promote gastric cancer cells invasion through a NF-kappa B and COX-2-mediated pathway

    Chun-Ying Wu; Chau-Jong Wang; Chi-Chuan Tseng; Hsiao-Ping Chen; Ming-Shing Wu; Jaw-Town Lin; Hiroyasu Inoue; Gran-Hum Chen

    AIM: To examine the effects of Helicobacter pylori (H pylori) infection on the invasiveness of gastric cancer cells, and to elucidate its mechanism. METHODS: Gastric carcinoma cells, MKN-45, were incubated with CagA-positive H pylori, and cell invasion was determined by Matrigel analysis. The expression of matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), and cyclooxygenase-2 (COX-2) were assessed by Western-blot analysis, and transcriptional activation of the COX-2 promoter was examined by measuring luciferase and beta-galactosidase activities. Lastly, the protein-DNA interaction was confirmed by an electrophoretic mobility shift assay. RESULTS: The current studies showed that: (1) incubation of CagA-positive H pylori with MKN-45 cells significantly promotes gastric cancer cells invasion, and this effect is attenuated by pre-treatment with NS-398, a COX-2 inhibitor, or PDTC, a nuclear factor kappa B (NF-kappa B) inhibitor; (2) the induction of MKN-45 cells invasion by H pylori is associated with increases in COX-2, MMP-9, and VEGF protein expression, and co-incubation of NS-398 or PDTC significantly reduces these effects; (3) H pylori infection transactivates COX-2 promoter activity and increases the binding of NF-kappa B to this promoter. CONCLUSION: Our data demonstrate that H pylori infection promotes gastric epithelial cells invasion by activating MMP-9 and VEGF expression. These effects appear to be mediated through a NF-kappa B and COX-2 mediated pathway, as COX-2 or NF-kappa B inhibitor significantly attenuate the invasiveness of gastric cancer cells and the expressions of MMP-9 and VEGF protein. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved., 2005年06月, WORLD JOURNAL OF GASTROENTEROLOGY, 11 (21), 3197 - 3203, web_of_science

    研究論文(学術雑誌)

  • C/EBP beta and its binding element are required for NF kappa B-induced COX2 expression following hypertonic stress

    J Chen; M Zhao; R Rao; H Inoue; CM Hao

    NF kappa B plays a critical role mediating COX2 expression in renal medullary interstitial cells (RMICs). The trans-activating ability of NF kappa B can be modified by another nuclear factor C/EBP beta that can physically bind to NF kappa B and regulate its activity. Because the COX2 promoter also contains a C/EBP beta site adjacent to the NF kappa B site, the present study examined whether these two transcription factors cooperate to induce COX2 expression following hypertonic stress. Hypertonicity markedly induced COX2 expression in cultured medullary interstitial cells by immunoblot analysis. The tonicity-induced COX2 expression was suppressed by mutant I kappa B (I kappa Bm) that blocks NF kappa B activation, demonstrating that tonicity-induced COX2 expression depends on NF kappa B activation. However, mutation of the NF kappa B site in the COX2 promoter failed to abolish tonicity-induced COX2 reporter activity. I kappa B kinase-1 (IKK1) significantly induced COX2-luciferase activity by 2.3-fold ( n = 10, p < 0.01); mutation of the NF kappa B site also failed to abolish IKK1-stimulated COX2 reporter activity ( 86 +/- 3.1% of wild type, p > 0.05, n = 4). Interestingly, mutation of the C/EBP beta site of the COX2 gene significantly reduced both IKK1 and hypertonicity-induced COX2 reporter activity ( p < 0.01). To further examine the potential role of C/EBP beta in tonicity-induced COX2 expression, a dominant negative C/EBP beta-p20 was transduced into RMICs. C/EBP beta-p20 markedly suppressed hypertonic ( 550 mOsm) induction of COX2 ( immunoblot) to a similar extent as I kappa Bm. No additional suppression was observed when both NF kappa B and C/EBP beta were simultaneously blocked by I kappa Bm and C/EBP beta-p20. Interestingly, IKK-induced COX2 expression was not only blocked by I kappa Bm, but also completely abolished by C/EBP beta-p20. Further studies demonstrated physical association of C/EBP beta to NF kappa B p65 by coimmunoprecipitation. Importantly, this interaction between C/EBP beta and NF kappa B was greatly enhanced following hypertonic stress. These studies indicate C/EBP beta is required for the transcriptional activation of COX2 by NF kappa B, suggesting a dominant role for the C/EBP beta pathway in regulating induction of RMIC COX2 by hypertonicity., 2005年04月, JOURNAL OF BIOLOGICAL CHEMISTRY, 280 (16), 16354 - 16359, doi;web_of_science

    研究論文(学術雑誌)

  • Selenomethionine regulates cyclooxygenase-2 (COX-2) expression through nuclear factor-kappa B (NF-kappa B) in colon cancer cells

    DR Cherukuri; AC Goulet; H Inoue; MA Nelson

    Previously, we showed that selenomethionine (Se-Met) inhibits growth of colon cancer cells via suppressing COX-2 expression at both mRNA and protein level. However, the molecular mechanism by which Se-Met suppresses COX-2 expression remains to be elucidated. To this end, we transiently transfected HCA-7 cells with different COX-2 promoter constructs followed by Se-Met treatment (90 mu M) for 12 h. The results suggested the role of nuclear factor-kappa B (NF-kappa B) in transcriptional regulation of COX-2. We also observed complete inhibition of DNA binding activity of NF-kappa B in Se-Met (90 mu M) treated HCA-7 cells as shown by electrophoretic mobility shift assay (EMSA). Supershift assays with anti-p65 antibody identified p65 subunit in the protein complex. We further demonstrate dose-dependent inhibition of nuclear translocation of NF-kappa B/p65 in Se-Met treated HCA-7 cells, which could explain the observed reduction in DNA binding of NF-kappa B/p65. These results suggest that Se-Met regulates COX-2 at transcriptional level by modulating the activity of NF-kappa B transcription factor., 2005年02月, CANCER BIOLOGY & THERAPY, 4 (2), 175 - 180, web_of_science

    研究論文(学術雑誌)

  • Chrysin suppresses lipopolysaccharide-induced cyclooxygenase-2 expression through the inhibition of nuclear factor for IL-6 (NF-IL6) DNA-binding activity

    KJ Woo; YJ Jeong; H Inoue; JW Park; TK Kwon

    Chrysin is a natural, biologically active compound extracted from many plants, honey and propolis. It possesses potent anti-inflammation, anti-cancer and anti-oxidation properties. The mechanism by which chrysin suppresses COX-2 expression remains poorly understood. In the present report, we investigated the effect of chrysin on the expression of COX-2 in lipopolysaccharide (LPS)-activated Raw 264.7 cells. Chrysin significantly suppressed the LPS-induced COX-2 protein and mRNA expression in a dose-dependent manner. The ability of chrysin to suppress the expression of the COX-2 was investigated using luciferase reporters controlled by various cis-elements in COX-2 promoter region. Mutational analysis and electrophoretic mobility shift assay verified that nuclear factor for IL-6 was identified as responsible for the chrysin-mediated COX-2 downregulation. These results will provide new insights into the anti-inflammatory and anti-carcinogenic properties of chrysin. (C) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved., 2005年01月, FEBS LETTERS, 579 (3), 705 - 711, doi;web_of_science

    研究論文(学術雑誌)

  • Induction of cyclooxygenase-2 overexpression in human gastric epithelial cells by Helicobacter pylori involves TLR2/TLR9 and c-Src-dependent nuclear factor-kappa B activation

    YJ Chang; MS Wu; JT Lin; BS Sheu; T Muta; H Inoue; CC Chen

    Gastric epithelial cells were incubated with a panel of clinical isolates of Helicobacter pylori, including nonulcer dyspepsia with gastritis (HS, n=20), gastric ulcer (HU, n=20), duodenal ulcer (HD, n=21), and gastric cancer (HC, n=20). HC strains induced a higher cyclooxygenase-2 (COX-2) expression than those from HS, HD, and HU. The bacterial virulence factors and the host cellular pathways were investigated. Virulence genes of iceA, vacA, babA2, cagA 3' repeat region, and hrgA failed to show any association with the disease status and COX-2 expression. Methylation-specific polymerase chain reaction revealed HC strains not affecting the methylation status of COX-2 promoter. Nuclear factor (NF)-kappaB, NF-interleukin 6, and cAMP response element were found to be involved in COX-2 induction. We explored a novel NF-kappaB activation pathway. The mutants of TLR2 and TLR9, but not TLR4, inhibited H. pylori-induced COX-2 promoter activity, and neutralizing antibodies for TLR2 and TLR9 abolished H. pylori-induced COX-2 expression. Phosphatidylinositol-specific phospholipase C (PI-PLC), protein kinase C (PKC), and Src inhibitors inhibited COX-2 induction. The dominant-negative mutants of NIK and various IkappaB kinase complexes, including IKKbeta (Y188F), IKKbeta (Y199F), and IKKbeta (FF), inhibited the COX-2 promoter activity. Phosphorylation of GST-IKKbeta (132-206) at Tyr(188) and Tyr(199) by c-Src was found after H. pylori infection. In summary, H. pylori induces COX-2 expression via activations of NF-kappaB, NF-interleukin 6, the cAMP response element. In NF-kappaB activation, H. pylori acts through TLR2/TLR9 to activate both the cascade of PI-PLCgamma/PKCalpha/c-Src/IKKalpha/beta and the cascade of NIK/IKKalpha/beta, resulting in the IkappaBalpha degradation and the expression of COX-2 gene. The COX-2 overexpression may contribute to the carcinogenesis in patients colonized with these strains., 2004年12月, MOLECULAR PHARMACOLOGY, 66 (6), 1465 - 1477, doi;web_of_science

    研究論文(学術雑誌)

  • Bradykinin B2 receptor mediates NF-kappa B activation and cyclooxygenase-2 expression via the Ras/Raf-1/ERK pathway in human airway epithelial cells

    BC Chen; CC Yu; HC Lei; MS Chang; MJ Hsu; CL Huang; MC Chen; Sheu, JR; TF Chen; TL Chen; H Inoue; CH Lin

    In this study, we investigated the signaling pathways involved in bradykinin (BK)-induced NF-kappaB activation and cyclooxygenase-2 (COX-2) expression in human airway epithelial cells (A549). BK caused concentration- and time-dependent increase in COX-2 expression, which was attenuated by a selective B2 BK receptor antagonist (HOE140), a Ras inhibitor (manumycin A), a Raf-1 inhibitor (GW 5074), a MEK inhibitor (PD 098059), an NF-kappaB inhibitor (pyrrolidine dithiocarbate), and an IkappaB protease inhibitor (L-1-tosylamido-2-phenylethyl chloromethyl ketone). The B1 BK receptor antagonist (Lys-(Leu(8))des-Arg(9)-BK) had no effect on COX-2 induction by BK. BK-induced increase in COX-2-luciferase activity was inhibited by cells transfected with the kappaB site deletion of COX-2 construct. BK-induced Ras activation was inhibited by manumycin A. Raf-1 phosphorylation at Ser(338) by BK was inhibited by manumycin A and GW 5074. BK-induced ERK activation was inhibited by HOE140, manumycin A, GW 5074, and PD 098059. Stimulation of cells with BK activated IkappaB kinase alphabeta (IKKalphabeta), IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 and p50 translocation from the cytosol to the nucleus, the formation of an NF-kappaB-specific DNA-protein complex, and kappaB-luciferase activity. BK-mediated increase in IKKalphabeta activity and formation of the NF-kappaB-specific DNA-protein complex were inhibited by HOE140, a Ras dominant-negative mutant (RasN17), manumycin A, GW 5074, and PD 098059. Our results demonstrated for the first time that BK, acting through B2 BK receptor, induces activation of the Ras/Raf-1/ERK pathway, which in turn initiates IKKalphabeta and NF-kappaB activation, and ultimately induces COX-2 expression inhuman airway epithelial cell line (A549)., 2004年10月, JOURNAL OF IMMUNOLOGY, 173 (8), 5219 - 5228, doi;web_of_science

    研究論文(学術雑誌)

  • Native LDL and oxidized LDL modulate cyclooxygenase-2 expression in HUVECs through a p38-MAPK, NF-kappa B, CRE dependent pathway and affect PGE(2) synthesis

    GD Norata; A Pirillo; F Pellegatta; H Inoue; AL Catapano

    Native low density lipoproteins (n-LDL) and oxidized low density lipoproteins (Ox-LDL) play a central role in atherogenesis and possess a wide variety of biological properties. We investigated whether n-LDL or Ox-LDL modulate cyclooxygenase-1 and -2 (Cox-1 and Cox-2) expression and prostaglandins release in human endothelial cells via an MAPK-dependent pathway. HUVECs were incubated in the presence of n-LDL or Ox-LDL (30 mug/ml for both) for 2-15 h. Real-time PCR, western blotting and immunocytochemistry were used to investigate Cox-1 and Cox-2 expression. N-LDL and Ox-LDL induced Cox-2 expression in a time- and dose-dependent manner. The Cox-2 protein was strongly induced 2 h after exposure to n-LDL or Ox-LDL, the induction was maximal after 4 h and sustained for at least 8 h. The effect was specific for Cox-2, as Cox-1 expression was not modulated either by n-LDL or by Ox-LDL. The induction of Cox-2 expression was mainly dependent on the activation of p38 MAPK. Transient transfection analysis using a Cox-2 promoter showed that n-LDL and Ox-LDL exert their effects at the transcriptional level via NF-kappaB and CREB activation. N-LDL and Ox-LDL increased PGE(2) release in a Cox-2-dependent manner while TXA(2) and PGI(2) release were not affected either by n-LDL or Ox-LDL. The finding that n-LDL and Ox-LDL induces Cox-2 in human endothelial cells through a p38 MAPK, NF-kappaB, CREB dependent pathway thus modulating PGE2 release, suggests a new mechanism by which these lipoproteins induce endothelial dysfunction, sustaining inflammatory processes in the arterial wall., 2004年09月, INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, 14 (3), 353 - 359, web_of_science

    研究論文(学術雑誌)

  • gamma-Mangostin inhibits inhibitor-kappaB kinase activity and decreases lipopolysaccharide-induced cyclooxygenase-2 gene expression in C6 rat glioma cells.

    Keigo Nakatani; Tohru Yamakuni; Nobuhiko Kondo; Tsutomu Arakawa; Kenji Oosawa; Susumu Shimura; Hiroyasu Inoue; Yasushi Ohizumi

    We investigated the effect of gamma-mangostin purified from the fruit hull of the medicinal plant Garcinia mangostana on spontaneous prostaglandin E(2) (PGE(2)) genase release and inducible cyclooxy-2 (COX-2) gene expression in C6 rat glioma cells. An 18-h treatment with gamma-mangostin potently inhibited spontaneous PGE(2) release in a concentration-dependent manner with the IC(50) value of approximately 2 microM, without affecting the cell viability even at 30 microM. By immunoblotting and reverse-transcription polymerase chain reaction, we showed that gamma-mangostin concentration-dependently inhibited lipopolysaccharide (LPS)-induced expression of COX-2 protein and its mRNA, but not those of constitutive COX-1 cyclooxygenase. Because LPS is known to stimulate inhibitor kappaB (IkappaB) kinase (IKK)-mediated phosphorylation of IkappaB followed by its degradation, which in turn induces nuclear factor (NF)-kappaB nuclear translocation leading to transcriptional activation of COX-2 gene, the effect of gamma-mangostin on the IKK/IkappaB cascade controlling the NF-kappaB activation was examined. An in vitro IKK assay using IKK protein immunoprecipitated from C6 cell extract showed that this compound inhibited IKK activity in a concentration-dependent manner, with the IC(50) value of approximately 10 microM. Consistently gamma-mangostin was also observed to decrease the LPS-induced IkappaB degradation and phosphorylation in a concentration-dependent manner, as assayed by immunoblotting. Furthermore, luciferase reporter assays showed that gamma-mangostin reduced the LPS-inducible activation of NF-kappaB-and human COX-2 gene promoter region-dependent transcription. gamma-Mangostin also inhibited rat carrageenan-induced paw edema. These results suggest that gamma-mangostin directly inhibits IKK activity and thereby prevents COX-2 gene transcription, an NF-kappaB target gene, probably to decrease the inflammatory agent-stimulated PGE(2) production in vivo, and is a new useful lead compound for anti-inflammatory drug development., 2004年09月, Molecular pharmacology, 66 (3), 667 - 74, 国際誌, pubmed

    研究論文(学術雑誌)

  • HDL3 induces cyclooxygenase-2 expression and prostacyclin release in human endothelial cells via a p38 MAPK/CRE-dependent pathway: Effects on COX-2/PGI-synthase coupling

    GD Norata; E Callegari; H Inoue; AL Catapano

    Objective - In endothelial cells, cyclooxygenase-1 (COX-1) and COX-2 both contribute to prostacyclin production. Recent findings suggest that COX-2 contributes significantly to systemic prostacyclin synthesis in humans; whether COX-2 inhibition is related to an increased cardiovascular risk is undergoing debate. HDLs have been shown to increase prostacyclin synthesis, thus in the present study we investigated the molecular mechanisms involved in this effect in endothelial cells. Methods and Results - HDL3 (30 mug/mL) induced COX-2 expression in a time- and dose-dependent manner. COX-2 was found mainly in the perinuclear area where it co-localizes with PGI synthase. Transient transfection experiments showed that CRE is required for HDL-induced COX-2 transcription, and we demonstrated that p38 MAPK activation by HDL3 is involved in COX-2 mRNA transcription and stabilization. As a consequence of COX-2-induction by HDL3 prostacyclin production increased, incubation with a COX-2 selective inhibitor blocked this effect. Moreover, HDL3 increased caveolin-1 phosphorylation, thus promoting PGI-synthase shuttling from the membrane to the perinuclear area. Conclusion - We conclude that in endothelial cells, HDL modulates COX-2/PGI-S activity via both p38 MAPK-dependent COX-2 mRNA stability and transcription and both caveolin-1 - dependent PGI-synthase shuttling and COX-2 coupling. The understanding of these mechanisms may provide new insights into the antiatherogenic role of HDL., 2004年05月, ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 24 (5), 871 - 877, doi;web_of_science

    研究論文(学術雑誌)

  • Involvement of the 3 '-untranslated region of cyclooxygenase-2 gene in its post-transcriptional regulation through the glucocorticoid receptor

    T Nishimori; H Inoue; Y Hirata

    Functional roles of the 3'-untranslated region (3'-UTR) of the human Cyclooxygenase-2 (COX-2) gene were evaluated by transient transfection using luciferase (Luc) reporter vectors into bovine arterial endothelial cells (BAEC). Insertion of the 3'-UTR into the downstream of a Luc coding region resulted in decreased reporter activity (23%), although insertion into the upstream was no effect. The reporter activity of the downstream insertion but not the upstream insertion was induced by bacterial lipopolysaccharide (LPS). Moreover, LPS selectively stabilized COX-2 mRNA. Next, to evaluate the role of the 3'-UTR together with glucocorticoid receptor (GR), a GR-expression vector was cotransfected with the reporter vector of the downstream insertion of the 3'-UTR. As a result, the LPS-induced reporter activity was suppressed by dexamethasone in a dose-dependent manner. These data suggest that the 3'-UTR of the COX-2 gene is involved in not only the induction by LPS but also the suppression by DEX of COX-2 expression at the post-transcriptional level. (C) 2004 Elsevier Inc. All rights reserved., 2004年04月, LIFE SCIENCES, 74 (20), 2505 - 2513, doi;web_of_science

    研究論文(学術雑誌)

  • Temporal and topographic profiles of cyclooxygenase-2 expression during 24 h of focal brain ischemia in rats

    C Yokota; T Kaji; Y Kuge; H Inoue; N Tamaki; K Minematsu

    Substantial increases in cyclooxygenase-2 (COX-2) mRNA and protein levels were demonstrated in the peri-infarct and focal ischemic areas after 3-24 and 12-24 h, respectively, in rats. In the ischemic core, significant increases in COX-2 mRNA followed 6 h of ischemia, though the peak level was about one-third of that in the peri-infarct area. Increases in COX-2 protein in the ischemic core were not observed during ischemic periods. Diffuse, neuronal COX-2 staining was found in peri-infarct areas as well as in discrete, immunoreactive neurons in the ischemic core. Robust increases in prostaglandin E-2 levels in the peri-infarct area were demonstrated following 24 h of ischemia. Prostaglandin production as well as COX-2 expression in ischemic tissues depended on the degree and duration of the reduction in cerebral blood flow. (C) 2003 Elsevier Ireland Ltd. All rights reserved., 2004年03月, NEUROSCIENCE LETTERS, 357 (3), 219 - 222, doi;web_of_science

    研究論文(学術雑誌)

  • Characterisation of [I-123]iomazenil distribution in a rat model of focal cerebral ischaemia in relation to histopathological findings

    T Kaji; Y Kuge; C Yokota; M Tagaya; H Inoue; T Shiga; K Minematsu; N Tamaki

    Iodine-123 labelled iomazenil ([I-123]IMZ) has been reported to be a useful marker of neuronal viability. The brain distribution of [I-123]IMZ, however, has not been correlated with the pathophysiological response in detail after an ischaemic insult. To characterise [I-123]IMZ as a marker of neuronal viability, we compared its brain distribution with cyclooxygenase-2 (COX-2) expression, DNA fragmentation and cellular integrity. [I-123]IMZ and [I-125]IMP were injected into rats with focal cerebral ischaemia for the purpose of dual-tracer autoradiography. COX-2 and microtubule-associated protein-2 (MAP-2, a marker of cellular integrity) were immunostained. In situ DNA polymerase-I-dependent dUTP incorporation into damaged DNA was used as an indicator of DNA fragmentation. Lesion to normal ratios (LNRs) for [I-123]IMP and [I-125]IMZ were calculated. [I-123]IMZ accumulation was preserved in several regions with impaired [I-123]IMP accumulation. COX-2 expression was occasionally observed, whereas neither DNA fragmentation nor MAP-2 denaturation was detected in these regions. DNA fragmentation and impaired MAP-2 immunostaining were observed only in the regions with reduced LNRs for both tracers. The LNR for [I-123]IMZ was significantly lower in regions with impaired MAP-2 immunostaining (0.120+/-0.152, P<0.0001), in regions positive for dUTP incorporation (0.488+/-0.166, P<0.0001) and in regions positive for COX-2 expression (0.626+/-0.186, P<0.001) than in histologically normal regions (0.784+/-0.213). Thus, neuronal DNA is still intact and cellular integrity is maintained in the ischaemic regions with preserved [I-123]IMZ accumulation. The impairment of [I-123]IMZ accumulation precedes DNA fragmentation and denaturation of cellular integrity. These results provide the molecular basis of [I-123]IMZ distribution., 2004年01月, EUROPEAN JOURNAL OF NUCLEAR MEDICINE AND MOLECULAR IMAGING, 31 (1), 64 - 70, doi;web_of_science

    研究論文(学術雑誌)

  • Tyrosine Phosphorylation of I-kappaB Kinase alpha/beta by Protein Kinase C-Dependent c-Src Activation Is Involved in TNF-alpha-Induced Cyclooxygenase-2 Expression.

    井上 裕康; W.C. Huang; J.J. Chen JJ; H. Inoue; C.C. Chen

    2004年, J. Immunol., 170, 4767 - 4775

  • Neuronal cyclooxygenase-2 expression during spreading depression and focal brain ischemia

    井上 裕康; Chiaki Yokota; Yuji Kuge; Yasuhiro Hasegawa; Hiroyasu Inoue; Masafumi Tagaya; Takeo Abumiya; Go Kito; Nagara Tamaki; Kazuo Minematsu

    2004年, 脳循環代謝, 16 (2), 89 - 95

  • Neuronal cyclooxygenase-2 induction associated with spreading depression and focal brain ischemia in primates

    C Yokota; Y Kuge; Y Hasegawa; H Inoue; M Tagaya; T Abumiya; G Kito; N Tamaki; K Minematsu

    We investigated pathophysiology of a primate model eliciting spreading depression (SD), as well as a primate thromboembolic model, by using PET. Immediately after the first SD, focal cortical hyperemia was demonstrated without being followed by spreading or persistent hypoperfusion. Cyclooxygenase-2 (COX-2) induction was detected in SD animals by microarray analysis. Immunoreactive neurons were observed in SD animals. In the thromboembolic model, cerebral blood flow (CBF) following 24 h of ischemia reduced to 20-40% in the ischemic temporal cortex as well as ischemic basal ganglia, while the reduction was 40-60% in the ischemic parietal cortex. Upregulation of COX-2 mRNA expression was observed after 2 h of ischemia, but disappeared by 24 h in the ischemic temporal cortex. In the ischemic parietal cortex, where CMRglc, vas preserved, COX-2 expression persisted even after 24 h of ischemia. In conclusion, we showed unique features of CBF changes associated with SD in primates. Neuronal COX-2 induction was demonstrated in SD animals as well as within potentially viable hypoperfused brain areas in primates. (C) 2004 Elsevier B.V. All rights reserved., 2004年, PET AND MOLECULAR IMAGING: STATE OF THE ART AND FUTURE PERSPECTIVES, 1264, 191 - 196, doi;web_of_science

    研究論文(国際会議プロシーディングス)

  • Characterization of [123I]iomazenil distribution in a rat model of focal cerebral ischemia in comparison with pathophysiological findings.

    Kaji T; Kuge Y; Yokota C; Tagaya M; Inoue H; Shiga T; Minematsu K; Tamaki N

    2004年, Eur J Nucl Med Mol Imaging., 31 (1), 64 - 70

  • Brain protection by resveratrol and fenofibrate against stroke requires peroxisome proliferator-activated receptor α in mice

    井上 裕康; H. Inoue; X. Jiang; T. Katayama; S. Osada; K. Umesono; S. Namura

    Peroxisome proliferator-activated receptors (PPARs) are ligand-dependent transcription factors which belong to the nuclear receptor family. We examined whether PPARalpha agonists and resveratrol, a polyphenol contained in grapes, protect the brain against ischemia. To investigate whether resveratrol activates PPARs, we performed a cell-based transfection activity assay using luciferase reporter plasmid. PPARalpha and PPARgamma were activated by resveratrol in primary cortical cultures and vascular endothelial cells. Resveratrol (20 mg/kg, 3 days) reduced infarct volume by 36% at 24 h after middle cerebral artery occlusion in wild-type mice. The PPARalpha agonists fenofibrate (30 mg/kg, 3 days) and Wy-14643 (30 mg/kg, 7 days) exerted similar brain protection. However, resveratrol and fenofibrate failed to protect the brain in PPARalpha knockout mice. The data indicate that PPARalpha agonists protect the brain through PPARalpha. (C) 2003 Elsevier Ireland Ltd. All rights reserved., 2003年12月, Neurosci. Lett, 352 (3), 203 - 206, doi;web_of_science

    研究論文(学術雑誌)

  • Transcriptional regulation of cyclooxygenase 2 by bradykinin and interleukin-1 beta in human airway smooth muscle cells: Involvement of different promoter elements, transcription factors, and histone H4 acetylation

    M Nie; LH Pang; H Inoue; AJ Knox

    Bradykinin and interleukin-1beta (IL-1beta) induce cyclooxygenase 2 (COX-2) in human airway smooth muscle cells. Here we extended our study to explore the gene transcriptional regulation. By transfection with various COX-2 promoter reporter constructs, we found that the bp -327-to-+59 promoter region was essential for COX-2 gene transcription by bradykinin and IL-1beta and that the cyclic AMP response element (CRE) was critical in bradykinin-induced transcription, whereas nuclear factor IL-6 and CRE and, to a lesser extent, nuclear factor-kappaB (NF-kappaB) were involved in IL-1beta-induced transcription. An electrophoretic mobility shift assay revealed that both bradykinin and IL-1beta elicited CRE-binding protein-1 (CREB-1) binding, and IL-1beta also elicited CCAAT/enhancer-binding protein beta and NF-kappaB binding to their respective elements in the COX-2 promoter. These transcription factors were associated with the COX-2 promoter, which was dynamically linked to different patterns of histone H4 acetylation by bradykinin and IL-1beta, as demonstrated by chromatin immunoprecipitation. We also revealed that endogenous prostaglandin E-2 was critical in bradykinin-induced COX-2 transcription initiation and involved in IL-1beta-induced COX-2 transcription at a latter stage. Our result provide the first evidence that COX-2 transcriptional regulation by different stimuli involves different promoter elements and transcription factors and is associated with chromatin remodeling after selective histone H4 acetylation in a stimullus-specific manner., 2003年12月, MOLECULAR AND CELLULAR BIOLOGY, 23 (24), 9233 - 9244, doi;web_of_science

    研究論文(学術雑誌)

  • Control of COX-2 gene expression through peroxisome proliferator-activated receptor gamma in human cervical cancer cells

    SW Han; H Inoue; LC Flowers; N Sidell

    Purpose: The peroxisome proliferator-activated receptor-gamma (PPARgamma), a ligand-dependent transcription factor belonging to the family of nuclear receptors, has been implicated in the control of cyclooxygenase (COX) 2 expression in some tissue, although the exact mechanism(s) of this activity has not been elucidated. In this study we explored the possible mechanism(s) of control of COX-2 gene expression through PPARgamma signaling in human cervical cancer. Experimental Design: Using primary human cervical tissues and the CaSki human cervical cancer cell line, we assayed for PPARgamma and COX-2 mRNA expression by reverse transcription-PCR. Nuclear protein binding activities to three response elements located in the COX-2 promoter [nuclear factor kappaB (NFkappaB), cyclic AMP response element, and activator protein (AP)-2] were measured by gel mobility shift assays. We used transient transfection assays with COX-2 promoter reporter gene constructs to determine the regulatory sites in this promoter, which mediates PPARgamma regulation of COX-2 activity. Results: We showed, for the first time, that primary human cervical cancer tissues express PPARgamma. Using CaSki cells, we demonstrated that COX-2 and PPARgamma mRNA levels were inversely regulated by PPARgamma ligands in that these compounds up-regulated PPARgamma but down-regulated COX-2. In contrast, epidermal growth factor (EGF), a potent activator of COX-2, decreased PPARgamma mRNA levels. This down-regulation of PPARgamma mRNA by EGF was blocked in the presence of NS-398, a selective COX-2 inhibitor. PPARgamma ligands suppressed the binding activities of AP-1 (binding to CRE) and NFkappaB but not AP-2. Transient transfection results indicated that EGF stimulated whereas PPARgamma ligands inhibited COX-2 promoter (-327/+59) activity. This effect by PPARgamma ligands on the COX-2 promoter was blocked when the CRE, but not the NFkappaB, binding site was mutagenized. Conlcusion: Cervical cancer cells express readily detectable levels of PPARgamma. There is reciprocal negative regulation between COX-2 and PPARgamma signaling in human cervical cancer cells. The ability of PPARgamma ligands to inhibit COX-2 appears to be mediated predominantly through inhibition of AP-1 protein binding to the CRE site in the COX-2 promoter., 2003年10月, CLINICAL CANCER RESEARCH, 9 (12), 4627 - 4635, web_of_science

    研究論文(学術雑誌)

  • 15-Deoxy-Δ12,14-prostaglandin J2 and laminar shear stress stabilize c-IAP1 in vascular endothelial cells.

    井上 裕康; Y. Taba; M. Miyagi M; Y. Miwa; H. Inoue; F. Takahashi-Yanaga; S. Morimoto; T. Sasaguri

    Laminar shear stress strongly inhibits vascular endothelial cell apoptosis by unknown mechanisms. We reported that shear stress stimulates endothelial cells to produce 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)) by elevating the expression level of lipocalin-type prostaglandin D synthase. To investigate the role of 15d-PGJ(2) produced in the vascular wall, we examined the effect of 15d-PGJ(2) on endothelial cell apoptosis. We induced apoptosis in human umbilical vein endothelial cells (HUVECs) by growth factor deprivation. 15d-PGJ(2) strongly inhibited DNA ladder formation, nuclear fragmentation, and caspase-3-like activity in HUVECs. To elucidate the mechanism by which 15d-PGJ(2) inhibits endothelial cell apoptosis, we examined expression of the inhibitor of apoptosis proteins (IAP) cellular-IAP1 (c-IAP1), c-IAP2, x-linked IAP, and survivin in HUVECs. In parallel with the inhibition of apoptosis, 15d-PGJ(2) elevated the expression level of c-IAP1 protein in a dose- and time-dependent manner without changing the mRNA level. Laminar shear stress also induced c-IAP1 expression. Chase experiments with the use of cycloheximide revealed that 15d-PGJ(2) and shear stress both inhibited the proteolytic degradation of c-IAP1 protein. These results suggested that 15d-PGJ(2) inhibits endothelial cell apoptosis through, at least in part, c-IAP1 protein stabilization. This mechanism might be involved in the antiapoptotic effect of laminar shear stress., 2003年07月, Am J Physiol Heart Circ Physiol., 285 (1), H38 - H46, doi;web_of_science

    研究論文(学術雑誌)

  • Cyclooxygenase-2 expression associated with spreading depression in a primate model

    C Yokota; H Inoue; Y Kuge; T Abumiya; M Tagaya; Y Hasegawa; N Ejima; N Tamaki; K Minematsu

    The authors previously provided evidence that spreading depression (SD) can be evoked in primates. Cyclooxygenase-2 (COX-2) expression has been found to increase in the rodent cortex undergoing SD, and the authors sought to determine whether this association exists in primate brain. In the present study, neuronal COX-2 expression was induced during SD in the primate cortex. The mean expression ratio of COX-2 messenger RNA in animals with SD was significantly higher than that measured in controls (1.69 vs. 0.5; P = 0.02). Induction of COX-2 in these animals was also detected by human microarray analysis. Results show that, as in rodents, neuronal COX-2 is induced in the primate cortex in response to SD., 2003年04月, JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM, 23 (4), 395 - 398, doi;web_of_science

    研究論文(学術雑誌)

  • Post-ischemic cyclooxygenase-2 expression is regulated by the extent of cerebral blood flow reduction in non-human primates

    C Yokota; Y Kuge; H Inoue; M Tagaya; G Kito; T Susumu; N Tamaki; K Minematsu

    We determined whether up to 24 h of ischemia could induce the expression of cyclooxygenase-2 (COX-2) in the brain of nonhuman primates. Randomized animals were subjected to either a 2 h ischemia (group II; n,= 3) or a 24 h ischemia (group 111; n = 3). Three animals in group I served as controls. In group 111, regional cerebral blood flow (CBF) and the cerebral glucose metabolic rate (CMRglc) were evaluated using positron emission tomography. Upregulation of COX-2 mRNA expression was observed after 2 h of ischemia, but disappeared by 24 h in the ischemic temporal cortex, in which both CMRglc and CBF were markedly reduced. In the ischemic parietal cortex, where CMRglc was preserved, COX-2 expression persisted even 24 h after ischemia. This study is the first to demonstrate neuronal COX-2 induction within potentially viable hypoperfused brain areas in nonhuman primates. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved., 2003年04月, NEUROSCIENCE LETTERS, 341 (1), 37 - 40, doi;web_of_science

    研究論文(学術雑誌)

  • Induction of cyclooxygenase-2 expression by fluid shear stress in vascular endothelial cells

    H Inoue; Y Taba; Y Miwa; C Yokota; M Miyagi; T Sasaguri

    2003年, ADVANCES IN PROSTAGLANDIN, LEUKOTRIENE, AND OTHER BIOACTIVE LIPID RESEARCH: BASIC SCIENCE AND CLINICAL APPLICATIONS, 525 (141-144), 141 - 144, web_of_science

    研究論文(学術雑誌)

  • Transcriptional and posttranscriptional regulation of cyclooxygenase-2 expression by fluid shear stress in vascular endothelial cells

    H Inoue; Y Taba; Y Miwa; C Yokota; M Miyagi; T Sasaguri

    Objective-Fluid shear stress induces cyclooxygenase (COX)-2 gene expression in vascular endothelial cells. We investigated the underlying mechanism of this induction. Methods and Results-Exposure of human umbilical vein endothelial cells to laminar shear stress in the physiological range (1 to -30 dyne/cm(2)) upregulated the expression of COX-2 but not COX-1, a constitutive isozyme of COX. The expression of COX-2 mRNA began to increase within 0.5 hour after the loading of shear stress and reached a maximal level at 4 hours. Roles of the promoter region and the 3'-untranslated region in the human COX-2 gene were evaluated by the transient transfection of luciferase reporter vectors into bovine arterial endothelial cells. Shear stress elevated luciferase activity via the region between -327 and 59 bp. Mutation analysis indicated that cAMP-responsive element (-59/-53 bp) was mainly involved in this response. On the other hand, shear stress selectively stabilized COX-2 mRNA. Moreover, shear stress elevated luciferase activity when a 3'-untranslated region of COX-2 gene containing 17 copies of the AUUUA mRNA instability motif was inserted into the vector. Conclusions-Transcriptional activation and posttranscriptional mRNA stabilization contribute to the rapid and sustained expression of COX-2 in response to shear stress., 2002年09月, ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY, 22 (9), 1415 - 1420, doi;web_of_science

    研究論文(学術雑誌)

  • Deoxycholic acid causes DNA damage in colonic cells with subsequent induction of caspases, COX-2 promoter activity and the transcription factors NF-kB and AP-1

    B Glinghammar; H Inoue; JJ Rafter

    Evidence is accumulating that bile acids induce apoptosis in colonic cells. Therefore, it becomes important to study the underlying molecular mechanisms and the role of this phenomenon in tumor promotion. Minutes after exposure of HCT 116 and HT-29 cells to deoxycholate (DCA), DNA damage, measured using the COMET assay, was evident. Caspase-3 was rapidly activated in HCT 116 cells exposed to DCA, whereas in HT-29 cells, caspase-3 activation was delayed. Using transient transfections with reporter constructs, we showed that the transcription factors activator protein-1 (AP-1) and NF-kB were increased in HCT 116 cells, in a dose-dependent fashion, by DCA COX-2 promoter activity was also induced by DC A and using mutant COX-2 promoter plasmids, we showed that the ability of DCA to induce promoter activity was partly dependent upon a functional NF-kB and C/EBP site, and completely dependent on a functional c-AMP response element site. DNA damage thus appears to be the initiating event in DCA-induced apoptosis. In conclusion, the bile acid, DCA, has a major impact on apoptotic mechanisms in colonic cells and this may be contributing to its effect as a tumor promoter., 2002年05月, CARCINOGENESIS, 23 (5), 839 - 845, web_of_science

    研究論文(学術雑誌)

  • Growth stimulation and epidermal growth factor receptor induction in cyclooxygenase-overexpressing human colon carcinoma cells

    T Yoshimoto; T Takahashi; T Kinoshita; T Sakashita; H Inoue; T Tanabe

    2002年, EICOSANOIDS AND OTHER BIOACTIVE LIPIDS IN CANCER, INFLAMMATION, AND RADIATION INJURY, 5, 507 (403-407), 403 - 407, web_of_science

    研究論文(学術雑誌)

  • Constitutive expression of the cyclooxygenase-2 gene in T-cell lines infected with human T cell leukemia virus type I

    N Mori; H Inoue; T Yoshida; T Tanabe; N Yamamoto

    Cyclooxygenase-2 (COX-2), an inducible enzyme that catalyzes the formation of prostaglandins and other eicosanoids from arachidonic acid, is constitutively expressed in several human carcinomas. COX-2 expression, however, has not been extensively studied in leukemia. Human T cell leukemia virus type I (HTLV-I) is the etiologic agent of adult T cell leukemia, an aggressive form of human T cell malignancy. We studied COX-2 mRNA expression in various human T-cell lines. Northern blot analysis revealed that COX-2 mRNA steady-state levels were high in 4 of 7 T-cell lines infected with HTLV-I. COX-2 mRNA, however, was not expressed in any of 3 HTLV-I-negative T-cell lines. We also confirmed COX-2 expression in 6 of 7 HTLV-I-positive T-cell lines by reverse transcription-PCR. HTLV-I Tax is known to increase the expression of cellular genes and thus we assayed Tax for its ability to increase transcription from the COX-2 promoter. Although Tax increased transcription of the COX-2 promoter in a T-cell line, Tax expression did not induce COX-2 mRNA expression, indicating that Tax alone is not sufficient for significant accumulation of COX-2 mRNA, which probably requires an additional viral protein. To evaluate the potential role of COX-2 in T cells, the HTLV-I-infected T-cell lines were treated with NS398, a selective COX-2 inhibitor. NS398 treatment inhibited proliferation and induced apoptosis of HTLV-I-infected T-cell lines and downregulated Bcl-2 and Bcl-X-L mRNA expression, followed by chromosomal DNA fragmentation. Our data suggest that COX-2 is expressed selectively in T-cell lines infected with HTLV-I and that this gene may play a role in cell survival. (C) 2001 Wiley-Liss, Inc., 2001年12月, INTERNATIONAL JOURNAL OF CANCER, 94 (6), 813 - 819, doi;web_of_science

    研究論文(学術雑誌)

  • Role of cyclic AMP responsive element in the UVB induction of cyclooxygenase-2 transcription in human keratinocytes

    QB Tang; WX Chen; MS Gonzales; J Finch; H Inoue; GT Bowden

    It has been shown that UVB irradiation induces expression of COX-2 and up-regulation of COX-2 plays a functional role in UVB tumor promotion. In this study, we examined the cis-elements in the human COX-2 promoter that may be responsible for the UVB induction of COX-2. Analyses with the COX-2 promoter region revealed that the cyclic AMP responsive element near the TATA box was essential for both basal and UVB induced COX-2 expression. This was further supported by studies using a dominant negative mutant of CREB, which strongly inhibited the activity of COX-2 promoter. Electrophoretic mobility shift assays indicated that CREB and ATF-I were the major proteins binding to the COX-2 CRE. CREB and ATF-I were phosphorylated upon UVB treatment, and SB202190, a p38 MAPK inhibitor, decreased the phosphorylation of CREB/ATF-1 and suppressed COX-2 promoter activity. In contrast, treatment with forskolin, an activator of adenylyl cyclase, led to phosphorylation of CREB and ATF-I and activation of COX-2 promoter. Finally, enhanced binding of phospho-CREB/ATF-1 to the COX-2 CRE was observed after UVB induction. Thus, one signaling pathway for UVB induction of human COX-2 involves activation of p38, subsequent phosphorylation of CREB/ATF-1, and activation of the COX-2 CRE through enhanced binding of phosphorylated CREB/ATF-1., 2001年08月, ONCOGENE, 20 (37), 5164 - 5172, doi;web_of_science

    研究論文(学術雑誌)

  • Transcriptional silencing of cyclooxygenase-2 by hyper-methylation of the 5 ' CpG island in human gastric carcinoma cells (vol 61, pg 4628, 2001)

    SH Song

    2001年08月, CANCER RESEARCH, 61 (15), 5956 - 5956, web_of_science

  • Roles of Akt and Glycogen Synthase Kinase 3 in the Ultraviolet B Induction of Cyclooxygenase-2 Transcription in Human Keratinocytes.

    井上 裕康; Q. Tang; M. Gonzales; H. Inoue; G. T. Bowden

    Ultraviolet B (UVB)-induced cyclooxygenase-2 (COX-2) expression plays an important role in WE tumor promotion. We examined whether Akt and glycogen synthase kinase 3 beta (GSK-3 beta), components of the phosphatidylinositol 3'-kinase pathway, are involved in UVB induction of COX-2 transcription. UVB caused Akt phosphorylation at both Thr-308 and Ser-473 that was inhibited by LY294002, a phosphatidylinositol 3'-kinase inhibitor. LY294002 also decreased the expression of endogenous COX-2 protein and a luciferase construct driven by COX-2 promoter. Similarly, UVB caused phosphorylation of GSK-3 beta (Ser-9) and presumably inactivation of GSK-3 beta. Inhibition of GSK-3 beta by lithium induced endogenous COX-2 protein expression and COX-2 promoter activity. Finally, overexpression of a dominant-negative Akt mutant or wild-type GSK-3 beta suppressed UVB-mediated induction of COX-2 promoter. These studies suggest that inactivation of GSK-3 beta through activation of Akt plays an important role in the UVB induction of COX-2 transcription., 2001年06月, Cancer Res., 61 (11), 4329 - 4332, web_of_science

    研究論文(学術雑誌)

  • Induction of cyclooxygenase-2 by Epstein-Barr virus latent membrane protein 1 is involved in vascular endothelial growth factor production in nasopharyngeal carcinoma cells

    S Murono; H Inoue; T Tanabe; Joab, I; T Yoshizaki; M Furukawa; JS Pagano

    Cyclooxygenase-2 (COX-2) is an inducible form of COX and is overexpressed in diverse tumors, raising the possibility of a role for COX-2 in carcinogenesis. In addition, COX-2 contributes to angiogenesis. The Epstein-Barr virus (EBV) oncoprotein, latent membrane protein 1 (LMP1), is detected in at least 70% of nasopharyngeal carcinoma (NPC) and all EBV-infected preinvasive nasopharyngeal lesions. We found that in specimens of LMP1-positive NPC, COX-2 is frequently expressed, whereas LMP1-negative NPC rarely express the enzyme. We next found that expression of LMP1 in EBV-negative nasopharyngeal epithelial cells induced COX-2 expression. Coexpression of I kappaB alpha (S32A/S36A). which is not phosphorylated and prevents NF-kappaB activation, with LMP1 showed that NF-kappaB is essential for induction of COX-2 by LMP1,We also demonstrate that NF-kappaB is involved in LMP1-induced cox-2 promoter activity with the use of reporter assays. Two major regions of LMP1, designated CTAR1 and CTAR2, are signal-transducing domains of LMP1, Constructs expressing either CTAR1 or CTAR2 induce COX-2 but to a lesser extent than wild-type LMP1, consistent with the ability of both regions to activate NF-kappaB, Furthermore, we demonstrate that LMP1-induced COX-2 is functional because LMP1 increased production of prostaglandin E-2 in a COX-2-dependent manner. Finally, we demonstrate that LMP1 increased production of vascular endothelial growth factor (VEGF), Treatment of LMP1-expressing cells with the COX-2-specific inhibitor (Ns-398) dramatically decreased production of VEGF, suggesting that LMP1-induced VEGF production is mediated, at least in part, by COX-2, These results suggest that COX-2 induction by LMP1 may play a role in angiogenesis in NPC., 2001年06月, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 98 (12), 6905 - 6910, doi;web_of_science

    研究論文(学術雑誌)

  • Reciprocal interactions between human T-lymphotropic virus type 1 and prostaglandins: Implications for viral transmission

    M Moriuchi; H Inoue; H Moriuchi

    Human T-lymphotropic virus type 1 (HTLV-1), the etiologic agent of adult T-cell leukemia/lymphoma, is transmitted through breast milk and seminal fluid, which are rich in prostaglandins (PGs), We demonstrate that PGE, upregulates the HTLV-1 long terminal repeat promoter through the protein kinase A pathway, induces replication of HTLV-1 in peripheral blood mononuclear cells (PBMC) derived from asymptomatic carriers, and enhances transmission of HTLV-1 to cord blood mononuclear cells (CBMC), Furthermore, HTLV-1 Tax transactivates a promoter for cyclooxygenase 2, a PG synthetase, and induces PGE(2) expression in PBMC or CBMC, Thus, HTLV-1 interacts with and benefits from PGs, constituents of its own vehicle for transmission., 2001年01月, JOURNAL OF VIROLOGY, 75 (1), 192 - 198, doi;web_of_science

    研究論文(学術雑誌)

  • 15-deoxy-Delta(12,14)-prostaglandin J(2) induces G(1) arrest and differentiation marker expression in vascular smooth muscle cells

    Y Miwa; T Sasaguri; H Inoue; Y Taba; A Ishida; T Abumiya

    In search of substances useful for the treatment of atherosclerotic vascular diseases, we studied the effects of 15-deoxy-(12,14)-prostaglandin J(2) (15d-PGJ(2)), a natural ligand for peroxisome proliferator-activated receptor gamma, on the proliferation and differentiation of vascular smooth muscle cells (VSMCs). 15d-PGJ(2) but not WY14643, an agonist for peroxisome proliferator-activated receptor alpha, dose-dependently inhibited VSMC proliferation; the effect was maximal at 12 mu M. This compound strongly suppressed the activities of cyclin-dependent kinases (Cdk) 4, 6, and 2, thereby preventing the phosphorylation of the retinoblastoma protein. These Cdks seemed to be inhibited through two mechanisms: the down-regulation of cyclin D1 and the up-regulation of Cdk inhibitor p21(Cip1/Waf1/Sdi1). 15d-PGJ(2) was found to inhibit the phosphatidylinositol 3-kinase/protein kinase B signaling pathway, which mediates cyclin D1 expression. Mitogenic stimulation of quiescent cells decreased the level of mRNA for the smooth muscle-specific myosin heavy-chain SM1, whereas this reduction was prevented by 15d-PGJ(2). A long-term treatment of exponentially growing VSMCs with 15d-PGJ(2) markedly elevated the mRNA level of SM1 and, moreover, induced SM2, another isoform expressed exclusively in mature VSMCs. 15d-PGJ(2) also increased the expression levels of calponin-h1 and smooth muscle alpha-actin. These results suggest that 15d-PGJ(2) induces G(1) arrest by two distinct mechanisms and promotes VSMC differentiation., 2000年10月, MOLECULAR PHARMACOLOGY, 58 (4), 837 - 844, web_of_science

    研究論文(学術雑誌)

  • Regulation of constitutive cyclooxygenase-2 expression in colon carcinoma cells

    JY Shao; HM Sheng; H Inoue; JD Morrow; RN DuBois

    Cyclooxygenase-2 (COX-2) is not normally expressed in the human large intestine, but its levels are increased in the majority of human colorectal carcinomas, Here we investigate the regulation of constitutive COX-2 expression and prostaglandin production in human colorectal carcinoma cells. Both COX-2 mRNA and protein were expressed in well differentiated HCA-7, Moser, LS-174, and HT-29 cells, albeit at different levels. COX-2 expression was not detected in several poorly differentiated colon cancer cell lines including DLD-1, Transcriptional regulation played a key role for the expression of COX-2 in human colon carcinoma cells, and both the nuclear factor for interleukin-6 regulatory element and the cAMP-response element were responsible for regulation of COX-2 transcription. COX-2 mRNA was more stable in HCA-7 cells than in the other cell lines tested. Both transcriptional and post-transcriptional regulation of COX-2 involved the MAP kinase pathway. Modulation of the Akt/protein kinase B or Rho B signaling pathways altered the levels of COX-2 expression. Furthermore, COX-2 protein is degraded through ubiquitin proteolysis, and its half-life was similar to3.5-8 h, HCA-7 cells produced significant quantities of prostaglandin E-2 and other prostaglandins. Moser and LS-174 cells also generated prostaglandins, but levels were significantly lower than that observed in HCA-7 cells., 2000年10月, JOURNAL OF BIOLOGICAL CHEMISTRY, 275 (43), 33951 - 33956, web_of_science

    研究論文(学術雑誌)

  • Feedback Control of COX-2 Expression through PPAR.

    井上 裕康; H. Inoue; T. Tanabe; K. Umesono

    Cyclooxygenase-2 (COX-2), a rate-limiting enzyme for prostaglandins (PG), plays a key role in inflammation, tumorigenesis, development, and circulatory homeostasis. The POD, metabolite 15-deoxy-Delta(12,14) PGJ(2) (15d-PGJ(2)) was identified as a potent natural ligand for the peroxisome proliferator-activated receptor-gamma (PPAR gamma), PPAR gamma expressed in macrophages has been postulated as a negative regulator of inflammation and a positive regulator of differentiation into foam cell associated with atherogenesis. Here, we show that 15d-PGJ(2) suppresses the Lipopolysaccharide (LPS)-induced expression of COX-2 in the macrophage-like differentiated U937 cells but not in vascular endothelial cells. PPAR gamma mRNA abundantly expressed in the U937 cells, not in the endothelial cells, is down-regulated by LPS, In contrast, LPS up-regulates mRNA for the glucocorticoid receptor which ligand anti-inflammatory steroid dexamethasone (DEX) strongly suppresses the LPS-induced expression of COX-2, although both 15d-PGJ(2) and DEX suppressed COX-2 promoter activity by interfering with the NF-kappa B signaling pathway, Transfection of a PPAR gamma expression vector into the endothelial cells acquires this suppressive regulation of COX-2 gene by 15d-PGJ(2) but not by DM. A selective COX-2 inhibitor, NS-398, inhibits production of PGD(2) in the U937 cells. Taking these findings together, we propose that expression of COX-2 is regulated by a negative feedback loop mediated through PPAR gamma, which makes possible a dynamic production of PC, especially in macrophages, and may be attributed to various expression patterns and physiological functions of COX-2., 2000年09月, J. Biol. Chem., 275 (36), 28028 - 28032, doi;web_of_science

    研究論文(学術雑誌)

  • Regulation of cyclooxygenase-2 by interferon gamma and transforming growth factor alpha in normal human epidermal keratinocytes and squamous carcinoma cells - Role of mitogen-activated protein kinases

    H Matsuura; M Sakaue; K Subbaramaiah; H Kamitani; TE Eling; AJ Dannenberg; T Tanabe; H Inoue; J Arata; AM Jetten

    Treatment of normal human epidermal keratinocytes (NHEK) with interferon-gamma (IFN-gamma) causes a 9-fold increase in the level of cyclooxggenase-2 (COX-2) mRNA expression. Nuclear run-off assays indicate that this induction is at least partly due to increased transcription. Activation of the epidermal growth factor receptor (EGFR) signaling pathway due to the enhanced transforming growth factor alpha (TGF alpha) expression plays an important role in the induction of COX-2 by IFN-gamma. This is supported by the ability of TGF alpha to rapidly induce COX-2 and the inhibition of the IFN-gamma-mediated COX-2 mRNA induction by an EGFR antibody and EGFR-selective kinase inhibitors. Deletion and mutation analysis indicates the importance of the proximal cAMP-response element/ATF site in the transcriptional control of this gene by TGF alpha. The increase in COX-2 mRNA by TGF alpha requires activation of both the extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways. Inhibition of p38 MAPK decreases the stability of COX-2 mRNA, while inhibition of MAPK/ERK kinase (MEK) does not. These results suggest that the p38 MAPK signaling pathway controls COX-2 at the level of mRNA stability, while the ERK signaling pathway regulates COX-2 at the level of transcription. In contrast to NHEK IFN-gamma and TGF alpha are not very effective in inducing TGF alpha or COX-2 expression in several squamous carcinoma cell lines, indicating alterations in both IFN-gamma and TGF alpha response pathways., 1999年10月, JOURNAL OF BIOLOGICAL CHEMISTRY, 274 (41), 29138 - 29148, doi;web_of_science

    研究論文(学術雑誌)

  • Activation of matrix metalloproteinase-2 in human breast cancer cells overexpressing cyclooxygenase-1 or-2

    Y Takahashi; F Kawahara; M Noguchi; K Miwa; H Sato; M Seiki; H Inoue; T Tanabe; T Yoshimoto

    Human breast cancer cell line Hs578T was stably transfected with cDNA for cyclooxygenase-1 or -2, When the cells overexpressing cyclooxygenase-1 or -2 mere stimulated with concanavalin A, the processing of matrix metalloproteinase-2 was observed with the aid of gelatin zymography, This processing was not seen in mock-transfected and original cells which did not express detectable cyclooxygenase activity. Furthermore, Northern blotting shelved 8-13 fold induction of membrane-type 1 matrix metalloproteinase,which processed matrix metalloproteinase-2 in the cells expressing cyclooxygenases. These findings suggest that both isoforms of cyclooxygenase mediate the processing of matrix metalloproteinase-2 through induction of membrane-type 1 metalloproteinase in breast cancer cells. (C) 1999 Federation of European Biochemical Societies., 1999年10月, FEBS LETTERS, 460 (1), 145 - 148, doi;web_of_science

    研究論文(学術雑誌)

  • Growth stimulation and induction of epidermal growth factor receptor by overexpression of cyclooxygenases 1 and 2 in human colon carcinoma cells

    T Kinoshita; Y Takahashi; T Sakashita; H Inoue; T Tanabe; T Yoshimoto

    There are two isoforms of cyclooxygenase (COX), COX-1 and COX-2. Recent epidemiological and experimental studies indicated a close relationship between COXs and the pathogenesis of colorectal cancer. The purpose of this study was to investigate the possible roles of both isoforms in the proliferation of colon carcinoma cells. A human colon carcinoma cell line, COLO 320DM, was transfected with an eukaryotic expression vector carrying cDNA of either COX-1 or COX-2, the expression of which was driven by a powerful elongation factor-la promoter in pEF-BOS. Both COX-1- and COX-2-expressing cells possessed a similar enzyme activity, 8-10 nmol/10 min per mg protein. Growth rates of both cell lines were stimulated by about 2-fold during a course of culture for 7 days as compared with mock-transfected cells. Although COX-1 and COX-2 are believed to have fundamentally different biological roles, essentially no differences in growth stimulation were observed between the COX-1 and COX-2 overexpressions in our experiments. The reason may be explained by high levels of COX expression, and subtle differences between the both cell lines would be possibly apparent by lower expression levels. The stimulated growth of the COX-transfected cells was accompanied by increased DNA synthesis as assessed by [H-3]thymidine incorporation. Furthermore, expression of epidermal growth factor receptor was markedly increased in these cells as examined by reverse transcription-polymerase chain reaction. A COX inhibitor, indomethacin, suppressed the stimulated growth, increased DNA synthesis and induction of epidermal growth factor receptor in COX-1- and COX-2-transfected cells. (C) 1999 Elsevier Science B.V. All rights reserved., 1999年04月, BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS, 1438 (1), 120 - 130, doi;web_of_science

    研究論文(学術雑誌)

  • Glucocorticoid-mediated suppression of the promoter activity of the cyclooxygenase-2 gene is modulated by expression of its receptor in vascular endothelial cells

    H Inoue; K Umesono; T Nishimori; Y Hirata; T Tanabe

    Cyclooxygenase-2 (COX-2), an inducible isozyme of cyclooxygenase, is expressed selectively in response to various inflammatory stimuli such as lipopolysaccharide (LPS) and its expression is suppressed by the glucocorticoid dexamethasone (DEX) in numerous types of cells. However, LPS-enhanced production of prostacyclin in bovine arterial endothelial cells (BAEC) was not significantly decreased by treatment with DEX but was suppressed by selective COX-2 inhibitors. This is consistent with the finding that DEX was not effective at preventing the expression of LPS-induced COX-2 mRNA. Transient transfection analysis showed that DEX did not suppress the LPS-induced promoter activity of the 5'-flanking region of the human COX-2 gene (nucleotides -327 to +59), Since RNA blot analysis indicated low-level expression of glucocorticoid receptor (GR) mRNA in BAEC, a GR-expression vector was transfected to evaluate the role of the GR in the COX-2 promoter activity. It was found that DEX mediated the suppression of the LPS-induced COX-2 promoter activity in a dose-dependent manner. These results suggest that the HEX-mediated suppression of LPS-induced promoter activity of the COX-2 gene is modulated by expression of the GR, which will be possible to account for a unique expression pattern of the COX-2 gene in BAEC. (C) 1999 Academic Press., 1999年01月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 254 (2), 292 - 298, doi;web_of_science

    研究論文(学術雑誌)

  • Localization of cyclooxygenase-2 and regulation of its mRNA expression in gastric ulcers in rats

    S Takahashi; JI Shigeta; H Inoue; T Tanabe; S Okabe

    It has been reported that cyclooxygenase-2 (COX-2) may play a crucial role in gastric ulcer healing. We examined the localization of COX-2 and the regulation of COX-2 mRNA expression in acetic acid ulcers in rats. PGE(2) production was elevated in ulcerated tissue but not in intact tissue. COX-2 mRNA expression was induced in only the ulcerated tissue, and COX-2 protein was found in fibroblasts, monocytes/macrophages, and granulocytes. A selective COX-2 inhibitor inhibited increased PGE(2) production by the ulcerated tissue. Interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta 1 (TGF-beta 1) mRNAs were also expressed only in the ulcerated tissue. In a culture of isolated ulcer base, blockade of IL-1 beta and TNF-alpha reduced COX-2 mRNA expression and PGE2 production. In contrast, COX-2 mRNA expression and PGE2 production were promoted by prevention of TGF-beta 1 action. These results indicate that COX-2 protein is highly localized in the base of gastric ulcers in rats and that COX-2 mRNA expression might be regulated positively by IL-1 beta and TNF-alpha and negatively by TGF-beta 1., 1998年11月, AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY, 275 (5), G1137 - G1145, web_of_science

    研究論文(学術雑誌)

  • Resveratrol inhibits cyclooxygenase-2 transcription and activity in phorbol ester-treated human mammary epithelial cells

    K Subbaramaiah; WJ Chung; P Michaluart; N Telang; T Tanabe; H Inoue; MS Jang; JM Pezzuto; AJ Dannenberg

    We determined whether resveratrol, a phenolic anti-oxidant found in grapes and other food products, inhibited phorbol ester (PMA)-mediated induction of COX-2 in human mammary and oral epithelial cells. Treatment of cells with PMA induces COX-2 and causes a marked increase in the production of prostaglandin E-2. These effects were inhibited by resveratrol, Resveratrol suppressed PMA-mediated increases in COX-2 mRNA and protein. Nuclear run-offs revealed increased rates of COX-2 transcription after treatment with PMA, an effect that was inhibited by resveratrol, PMA caused about a 6-fold increase in COX-2 promoter activity, which was suppressed by resveratrol, Transient transfections utilizing COX-2 promoter deletion constructs and COX-2 promoter constructs, in which specific enhancer elements were mutagenized, indicated that the effects of PMA and resveratrol were mediated via a cyclic AMP response element. Resveratrol inhibited PMA-mediated activation of protein kinase C, Overexpressing protein kinase C-alpha, ERK1, and c-Jun led to 4.7-, 5.1-, and C-fold increases in COX-2 promoter activity, respectively. These effects also were inhibited by resveratrol, Resveratrol blocked PMA-dependent activation of AP-l-mediated gene expression. In addition to the above effects on gene expression, we found that resveratrol also directly inhibited the activity of COX-2, These data are likely to be important for understanding the anti-cancer and anti-inflammatory properties of resveratrol., 1998年08月, JOURNAL OF BIOLOGICAL CHEMISTRY, 273 (34), 21875 - 21882, doi;web_of_science

    研究論文(学術雑誌)

  • Cloning and expression of the gene for a vanadium-dependent bromoperoxidase from a marine macro-alga, Corallina pilulifera

    M Shimonishi; S Kuwamoto; H Inoue; R Wever; T Ohshiro; Y Izumi; T Tanabe

    The cDNAs for a vanadium-dependent bromoperoxidase were cloned from a marine macro-alga, Covallina pilulifera. The open reading frame of one clone (bpo1) encoded a protein of 598 amino acids with a calculated molecular mass of 65312 Da in good agreement with that of 64 kDa determined for the native enzyme, The deduced amino acid sequence coincided web with partial sequences of peptide fragments of the enzyme. From the same cDNA library we also isolated another cDNA clone (bpo2) encoding a protein of 597 amino acids with an identity of about 90% to BPO1, suggesting a genetic diversity of the bromoperoxidase gene of C. pilulifera growing in a relatively narrow area. The carboxy-terminal 123 residues of the enzyme (BPO1) showed an identity of 45% to that of the marine macroalga Ascophillum nodosum. The homology search of the sequences of bromoperoxidases from C, pilulifera (this study) and A. nodosum, and chloroperoxidase from the fungus Curvalaria inaequalis indicated highly conserved sequences PxYxSGHA and LxxxxAxxRxxxGxHxxxD. Furthermore, it was found that the histidine residue directly bound to vanadium, other residues building up the metal center and catalytic histidine residue forming the active site of the chloroperoxidase from C, inaequalis are conserved in the primary structure of the bromoperoxidase from C, pilulifera. The cloned bpo1 was introduced into Escherichia coli, and the expressed BPO1 was purified from the recombinant strain. The N-terminal amino acid sequence of the purified BPO1 was identical to the deduced sequence from the cDNA except the N-terminal methionine. (C) 1998 Federation of European Biochemical Societies., 1998年05月, FEBS LETTERS, 428 (1-2), 105 - 110, doi;web_of_science

    研究論文(学術雑誌)

  • Transcriptional role of the nuclear factor kappa B site in the induction by lipopolysaccharide and suppression by dexamethasone of cyclooxygenase-2 in U937 cells

    H Inoue; T Tanabe

    Cyclooxygenase-2 (COX-2), an inducible isozyme of cyclooxygenase, is selectively expressed in response to lipopolysaccharide (LPS) and its expression is suppressed by the glucocorticoid dexamethasone (DEX) in the monocytic differentiated U937 cells, However, COX-2 mRNA was not detected nor induced by LPS before the cells differentiated. To study the transcriptional role of the NF-kappa B site (nucleotides -223 to -214) in the COX-2 gene, the luciferase reporter vector driven by the COX-2 promoter region (nucleotides -327 to +59) mutated at both the cAMP response element and the NF-IL6 site was stably transfected into U937 cells. The substantial luciferase activity observed in the undifferentiated cells was not induced by LPS, However, after the cells had differentiated, luciferase activity was induced by LPS and its induction was suppressed by DEX. Moreover, a protein tyrosine kinase inhibitor herbimycin A suppressed both the expression of COX-2 mRNA and the luciferase activity induced by LPS, These results suggest that the NF-kappa B site is involved in both the LPS-induced expression of the COX-2 gene and its suppression by DEX and herbimycin A in a differentiation-dependent manner. (C) 1998 Academic Press., 1998年03月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 244 (1), 143 - 148, doi;web_of_science

    研究論文(学術雑誌)

  • Cyclooxygenase expression in bovine aortic endothelial cells exposed to cyclic strain

    H Kito; C Yokoyama; H Inoue; T Tanabe; N Nakajima; BE Sumpio

    The aim of this study was to investigate the effect of cyclic strain on cyclooxygenase (COX)-1 and 2 expression in bovine aortic endothelial cells (EC). EC, subjected to 10% average strain at 60 cycle/min, were analyzed for induction of COX by Northern blot analysis and confirmed by analysis of promoter activity in transient transfection experiments. Exposure of EC to cyclic strain induced promoter activity and expression of COX-2 but not of COX-I. The extent of induction, however, was lower than that seen with stimulation of 12-O-tetradecanoylphorbol-13-acetate (TPA) and/or lipopolysaccharide (LPS). These results demonstrate that, unlike shear stress, cyclic strain does not affect COX-1 expression and is a weak inducer of COX-2 promoter activity in bovine aortic EC with minimal effect on mRNA expression., 1998年, ENDOTHELIUM-NEW YORK, 6 (2), 107 - 112, web_of_science

    研究論文(学術雑誌)

  • Retinoids suppress epidermal growth factor-induced transcription of cyclooxygenase-2 in human oral squamous carcinoma cells

    Mestre, JR; K Subbaramaiah; PG Sacks; SP Schantz; T Tanabe; H Inoue; AJ Dannenberg

    Cyclooxygenase-2 (Cox-2), the inducible form of cyclooxygenase, is up-regulated in tumors and transformed cells. Because this enzyme catalyzes the formation of prostaglandins from arachidonic acid, chemopreventive strategies that suppress its expression could be useful for preventing cancer. We investigated whether retinoids suppressed basal expression of Cox-2 or EGF-mediated induction of Cox-2 in human oral squamous carcinoma cells. Treatment with retinoids [all-trans-retinoic acid (all-trans-RA), 9-cis-RA, 13-cis-RA, and retinyl acetate] suppressed both basal levels of Cox-2 and EGF-mediated induction of Cox-2 protein and synthesis of prostaglandin E-2. Retinoids also suppressed the induction of Cox-2 mRNA by EGF, Transient transfection experiments showed that EGF caused about a 100% increase in Cox-2 promoter activity, an effect that was suppressed by retinoids. Levels of epidermal growth factor receptor were unaffected by retinoids. Epidermal growth factor caused a nearly 10-fold increase in mitogen-activated protein kinase activity; this effect was not blocked by retinoids., 1997年07月, CANCER RESEARCH, 57 (14), 2890 - 2895, web_of_science

    研究論文(学術雑誌)

  • Inverse gene expression of prostacyclin and thromboxane synthases in resident and activated peritoneal macrophages

    S Kuwamoto; H Inoue; Y Tone; Y Izumi; T Tanabe

    Prostacyclin and thromboxane A(2) produced from prostaglandin Hz are known to be important modulators with opposite biological activities. To examine possible roles of these prostanoids in immune responses, we have studied the gene expression of prostacyclin synthase (PGIS) and thromboxane synthase (TXS) in murine resident macrophages or in macrophages elicited with casein or bacillus Calmette-Gauerin (BCG). Northern blot analyses showed that the PGIS mRNA was expressed in a decreasing order in the resident, and casein- and BCG-elicited macrophages. In contrast, the TXS mRNA was expressed in an increasing order in the resident, and casein- and BCG-elicited macrophages. On the other hand, the mRNA for cyclooxygenase-2, which produces PGH(2) and participates in the production of prostanoids in inflammation, was expressed in both the resident and BCG-elicited macrophages but barely in the casein-elicited cells, In situ hybridization analysis showed that the expression of mRNAs for PGIS and TXS was ascribable not only to the alteration of the expression levels of both mRNAs in the each macrophage but also to the changes in subpopulations of the cells expressing these mRNAs. These observations suggested that the inverse gene expression of PGIS and TXS in macrophages contributes to immune responses by modulating the relative levels of prostacyclin and thromboxane A(2). (C) 1997 Federation of European Biochemical Societies., 1997年06月, FEBS LETTERS, 409 (2), 242 - 246, web_of_science

    研究論文(学術雑誌)

  • Benzo[a]pyrene up-regulates cyclooxygenase-2 gene expression in oral epithelial cells

    DJ Kelley; Mestre, JR; K Subbaramaiah; PG Sacks; SP Schantz; T Tanabe; H Inoue; JT Ramonetti; AJ Dannenberg

    Cyclooxygenase may be important in the pathogenesis of smoking-related cancer because it activates carcinogens and catalyzes prostaglandin biosynthesis. We determined the effects of benzo[a]pyrene (B[a]P), a polycyclic aromatic hydrocarbon in tobacco smoke, on cyclooxygenase-2 (Cox-2) mRNA, protein and synthesis of prostaglandin E(2) (PGE(2)) in normal and transformed oral epithelial cells. Treatment with B[a]P caused a dose-dependent increase in production of PGE(2), with a maximal increase of similar to 100%. Enhanced synthesis of PGE(2) was associated with increased amounts of Cox-2 protein. B[a]P also caused a two-fold increase in Cox-2 mRNA in both normal and transformed cells. Transient transfections with a Cox-2 promoter construct showed that B[a]P-mediated induction of Cox-2 mRNA reflected increased transcription, Levels of Cox-1 were unaffected by B[a]P, B[e]P did not affect the synthesis of PGE(2) or amounts of Cox-a, These data are important because B[a]P-mediated induction of Cox-2 may predispose to carcinogenesis by enhancing the production of mutagens and the synthesis of prostaglandins., 1997年04月, CARCINOGENESIS, 18 (4), 795 - 799, doi;web_of_science

    研究論文(学術雑誌)

  • Retinoids suppress phorbol ester-mediated induction of cyclooxygenase-2

    Mestre, JR; K Subbaramaiah; PG Sacks; SP Schantz; T Tanabe; H Inoue; AJ Dannenberg

    Cyclooxygenase-2 expression is up-regulated in transformed cells and tumors. Because this enzyme catalyzes the synthesis of prostaglandins, strategies aimed at suppressing its expression may prove useful in preventing or treating cancer, We investigated the ability of retinoids to suppress phorbol ester-mediated induction of cyclooxygenase-2 in human oral epithelial cells. Treatment with phorbol myristate acetate (PMA) resulted in approximately a 3-fold increase in the production of prostaglandin E(2) (PGE(2)). Retinoids [all-trans-retinoic acid (RA), 13-cis-RA, and retinyl acetate] markedly suppressed PMA-mediated increases in amounts of cyclooxygenase-2 (Cox-2) and the production of PGE(2). Retinoids also suppressed the induction of Cox-2 mRNA by PMA. Nuclear run-offs revealed increased rates of Cox-2 transcription after treatment with PMA; this effect was inhibited by all-trans-RA. Transient transfection experiments showed that PMA caused about a 2-fold increase in Cox-2 promoter activity, an effect that was suppressed by all-trans-RA. Our data indicate that treatment of oral epithelial cells with PMA is associated with enhanced transcription of Cox-2 and increased production of PGE(2). These effects of PMA were inhibited by retinoids., 1997年03月, CANCER RESEARCH, 57 (6), 1081 - 1085, web_of_science

    研究論文(学術雑誌)

  • The regional distribution and cellular localization of mRNA encoding rat prostacyclin synthase

    Yoshinori Tone; Hiroyasu Inoue; Shuntaro Hara; Chieko Yokoyama; Toshihisa Hatae; Hiroji Oida; Shu Narumiya; Ryuichi Shigemoto; Susumu Yukawa; Tadashi Tanabe

    The cloned cDNA for rat prostacyclin synthase was found to contain a 1503-bp open reading frame which encoded a 501-amino acid protein sharing 84% identity with the human enzyme. RNA blot analysis revealed that the rat prostacyclin synthase mRNA, as a single species of 2.1 kb, is expressed abundantly in the aorta and uterus. High levels of expression were also observed in the stomach, lung, heart, testis, liver, and skeletal muscle. Low but significant expression was also seen in the brain and kidney. Furthermore, the regional distribution and cellular localization of prostacyclin synthase mRNA were examined by in situ hybridization analysis of rat tissue sections. The definitive signals for the mRNA were localized in smooth muscle cells of the arteries, bronchi and uterus, and in the cells of the fibrous tunic surrounding the seminiferous tubules, which are characterized as smooth muscle cells. Besides smooth muscle cells, signal were also detected in the fibroblasts of the heart myocardium, lung parenchyma cells and kidney inner medulla tubules and interstitial cells., 1997年, European Journal of Cell Biology, 72 (3), 268 - 277, pubmed

    研究論文(学術雑誌)

  • Transcriptional regulation of human prostaglandin-endoperoxide synthase-2 gene in vascular endothelial cells

    H Inoue; T Tanabe

    1997年, EICOSANOIDS AND OTHER BIOACTIVE LIPIDS IN CANCER, INFLAMMATION, AND RADIATION INJURY 3, 407, 139 - 144, web_of_science

    研究論文(学術雑誌)

  • ヒト誘導型シクロオキシゲナーゼ遺伝子の構造と 血管内皮細胞におけるその発現誘導機構

    井上 裕康; 井上 裕康; 横山 知永子; 田辺 忠

    1997年, 血管, 20, 121 - 128

  • Human gene encoding prostacyclin synthase (PTGIS): Genomic organization, chromosomal localization, and promoter activity

    C Yokoyama; T Yabuki; H Inoue; Y Tone; S Hara; T Hatae; M Nagata; EI Takahashi; T Tanabe

    The prostacyclin synthase gene isolated from human genomic libraries (PTGIS) consists of 10 exons spanning approximately 60 kb. All the splice donor and acceptor sites conform to the GT/AG rule. Genomic Southern blot and fluorescence in situ hybridization analyses revealed that the human prostacyclin synthase gene is present as a single copy per haploid genome and is localized on chromosome 20q13.11-q13.13. The 1.5-kb sequence of the 5'-upstream of the translational initiation site contained both GC-rich and pyrimidine-rich regions and consensus sequences of the transcription factor recognition sites such as Sp1, AP-2, the interferon-gamma response element, GATA, NF-kappa B, the CACCC box, and the glucocorticoid response element. The core binding sequence (GAGACC) of the shear stress responsive element was also found in the 5'-flanking region of the gene. The major product of the primer extension analysis suggested that the transcription of the gene started from the positions around 49 bp upstream of the translational initiation codon. Transient transfection experiments using human aortic and bovine arterial endothelial cells demonstrated that the GC-rich region (positions -145 to -10) possessed a significant promoter activity. The 6-kb downstream sequence of the translational termination codon contained multiple polyadenylation signals, Alu repeat sequences, and the consensus sequence of the primate-repetitive DNA element, MER1. Two sizes of the prostacyclin synthase mRNAs (approximately 6 and 3.3 kb) were detected with the human aorta and lung. RNA blot hybridization analysis using the 3'-untranslated region as probe indicated that the sizes of the 3'-flanking regions were different in the major 6-kb and minor 3.3-kb mRNAs. (C) 1996 Academic Press, Inc., 1996年09月, GENOMICS, 36 (2), 296 - 304, doi;web_of_science

    研究論文(学術雑誌)

  • Site-directed mutagenesis of human prostacyclin synthase: Alteration of Cys(441) of the Cys-pocket, and Glu(347)and Arg(350) of the EXXR motif

    T Hatae; S Hara; C Yokoyama; T Yabuki; H Inoue; Ullrich, V; T Tanabe

    The possible active site Cys(441) in the Cys-pocket and Glu(347) and Arg(350) of the EXXR motif of the human prostacyclin synthase, which catalyzes the conversion of prostaglandin H-2 to prostacyclin, were subjected to site-directed mutagenesis in order to understand the role of these residues in expressing the enzymatic activity, Five expression vectors encoding the mutant enzymes with a single replacement, Cys(441)Ala, Cys(441)Ser, Cys(441)His, Glu(347)Ala and Arg(350)Ala, as well as the wild-type enzyme were expressed in 293 cells, The microsomal fraction of the cells expressing the wild-type enzyme showed a specific activity of 96 nmol 6-keto-PGF(1 alpha)/min per mg protein. All of the mutant enzymes examined showed no detectable enzyme activity, although immunoblot analysis demonstrated that levels of all the expressed mutant enzymes were similar to that of the wild-type enzyme, These results indicated that the Cys(441) in the Cys-pocket, and Glu(347) and Arg(350) of the EXXR motif of human prostacyclin synthase are important for expressing the enzymatic activity., 1996年07月, FEBS LETTERS, 389 (3), 268 - 272, doi;web_of_science

    研究論文(学術雑誌)

  • Protein tyrosine kinase inhibitors promote amylase secretion and inhibit ornithine decarboxylase induction in sialagogue-stimulated rat parotid explants

    F Kinoshita; A Ueno; Y Miwa; M Nishino; H Inoue

    Three sialagogues, isoproterenol (IPR), carbachol, and methoxamine, caused induction of ornithine decarboxylase (ODC) in cultured rat parotid explants. All the protein tyrosine kinase inhibitors tested suppressed this ODC induction but enhanced sialagogue-dependent amylase secretion. Sodium orthovanadate showed the reverse effects as the kinase inhibitors. Immunoblot analysis with anti-phosphotyrosine antibody revealed that herbimycin A depresses IPR-stimulated tyrosine phosphorylation of parotid proteins. Herbimycin A did not affect the IPR- or dibutyryl cAMP-induced surge of the parotid cAMP level but inhibited these agonist-dependent ODC inductions. These results suggest that sialagogue-induced ODC induction and amylase secretion are mediated by different signal transduction pathways and that protein tyrosine kinase participates in IPR-dependent ODC induction and amylase secretion in the process subsequent to the cAMP surge. (C) 1996 Academic Press, Inc., 1996年06月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 223 (1), 170 - 174, web_of_science

    研究論文(学術雑誌)

  • OVEREXPRESSION OF PROSTACYCLIN SYNTHASE INHIBITS GROWTH OF VASCULAR SMOOTH-MUSCLE CELLS

    S HARA; R MORISHITA; Y TONE; C YOKOYAMA; H INOUE; Y KANEDA; T OGIHARA; T TANABE

    To define the local effects of prostacyclin (PGI(2)) on the growth of vascular smooth muscle cells (VSMC), we transfected VSMC with an expression vector harboring the cDNA for PGI(2) synthase (PGIS), which catalyzes the rearrangement of prostaglandin H-2 to PGI(2). Transfection of the human PGIS cDNA into rat VSMC did not affect DNA synthesis under serum free basal conditions, but it increased PGI(2) synthesis and decreased DNA synthesis under serum-stimulated conditions (in the presence of 1 or 5% fetal calf serum). These results demonstrated that locally synthesized PGI(2) can exert autocrine and/or paracrine inhibitory effects on VSMC growth. It was also suggested that in vivo transfer of PGIS gene may be useful for the gene therapy for vascular disease such as neointimal hyperplasia. (C) 1995 Academic Press, Inc., 1995年11月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 216 (3), 862 - 867, doi;web_of_science

    研究論文(学術雑誌)

  • TRANSCRIPTIONAL REGULATION OF HUMAN PROSTAGLANDIN-ENDOPEROXIDE SYNTHASE-2 GENE BY LIPOPOLYSACCHARIDE AND PHORBOL ESTER IN VASCULAR ENDOTHELIAL-CELLS - INVOLVEMENT OF BOTH NUCLEAR FACTOR FOR INTERLEUKIN-6 EXPRESSION SITE AND CAMP RESPONSE ELEMENT

    H INOUE; C YOKOYAMA; S HARA; Y TONE; T TANABE

    There exist two distinct isozymes of prostaglandin-endoperoxide synthase (PES). PES-2 mRNA is synergistically induced by lipopolysaccharide (LPS) and 12-O-tetradecanoylphorbol-13-acetate (TPA) in bovine arterial endothelial cells. On the other hand, PES-1 mRNA is constitutively expressed under these conditions. Therefore, the promoter activities of the human genes for PES-1 and -2 in bovine arterial endothelial cells were examined. The 5'-flanking region of the human PES-2 gene (nucleotides -327 to +59) showed promoter activity inducible by LPS and TPA using transient transfection analysis, whereas that of the PES-1 gene (nucleotides -1010 to +69) showed constitutive promoter activity. Destruction of both consensus sequences for the nuclear factor responsible for the interleukin-6 expression (NF-IL6) site (nucleotides -132 to -124) and the cyclic AMP response element (CRE) (nucleotides -59 to -53) of the human PES-2 gene markedly reduced the promoter activity (25%) of the PES-2 gene after combined treatment with LPS and TPA, although single destruction of the NF-IL6 site or the CRE slightly reduced the promoter activity (60 or 90%, respectively). Moreover, cotransfection experiments showed that a trans-acting factor, CCAAT enhancer binding protein delta (C/EBP delta), which binds to both the NP-IL6 site and the CRE, increased the promoter activity of the PES-2 gene mainly through the CRE. C/EBP delta mRNA was rapidly induced by LPS. Collectively, these results suggest that transcription of the PES-2 gene in vascular endothelial cells is regulated through combination of the NF-IL6 site and the CRE and that C/EBP delta functions as one of the trans-acting factors., 1995年10月, JOURNAL OF BIOLOGICAL CHEMISTRY, 270 (42), 24965 - 24971, doi;web_of_science

    研究論文(学術雑誌)

  • REGULATION OF 2 ISOZYMES OF PROSTAGLANDIN ENDOPEROXIDE SYNTHASE AND THROMBOXANE SYNTHASE IN HUMAN MONOBLASTOID CELL-LINE U937

    T NANAYAMA; S HARA; H INOUE; C YOKOYAMA; T TANABE

    The mechanism responsible for the rapid increase of thromboxane A(2) synthesis by cells of the human monoblastoid cell line U937, which were differentiated with 12-O-tetradecanoyl-phorbol-13-acetate, induced by lipopolysaccharide (LPS) was studied. Both RNA blot and immunoblot analyses showed that LPS increased the levels of prostaglandin endoperoxide synthase-l (PES-I) and -2 (PES-2) in a time-dependent manner, and the modes of induction of the two isozymes differed. The maximum PES-1 mRNA level was 1.6 times higher 36 h after than before stimulation by LPS, and that of PES-P mRNA was elevated about 20-fold at its peak at 12 h after stimulation. Consequently, the immunoreactive PES-I and PES-L? protein levels also increased time-dependently after LPS stimulation. However, the effects of LPS on the thromboxane synthase mRNA and protein levels were much less marked. These results indicate that LPS-induced thromboxane synthesis by the differentiated cells was regulated at the levels of the two PES isozymes, predominantly at the PES-2 level., 1995年06月, PROSTAGLANDINS, 49 (6), 371 - 382, doi;web_of_science

    研究論文(学術雑誌)

  • EXPRESSION OF THE CHOLINE-ACETYLTRANSFERASE GENE DEPENDS ON PROTEIN KINASE-A ACTIVITY

    H INOUE; YP LI; JA WAGNER; LB HERSH

    Choline acetyltransferase activity is barely detectable in a mutant pheochromocytoma PC12 cell line, A123.7, which is deficient in protein kinase A activity. Northern blot and polymerase chain reaction analyses showed that this mutant cell line has dramatically reduced levels of choline acetyltransferase mRNA, which correlates with the low level of enzyme activity. Transient transfection analysis was used to assess the functionality, in these cells, of an enhancer element and a cholinergic-specific repressor element derived from the human choline acetyltransferase gene. The results show that the enhancer element is inactive in the protein kinase A-deficient cell line. Cotransfection experiments with plasmids expressing the catalytic subunit of protein kinase A support this conclusion. These data indicate that protein kinase A regulates expression of the choline acetyltransferase gene at the transcriptional level by controlling the activity of an enhancer element., 1995年03月, JOURNAL OF NEUROCHEMISTRY, 64 (3), 985 - 990, doi;web_of_science

    研究論文(学術雑誌)

  • STRUCTURE AND EXPRESSION OF THE HUMAN PROSTAGLANDIN ENDOPEROXIDE SYNTHASE-2 GENE

    H INOUE; T KOSAKA; A MIYATA; S HARA; C YOKOYAMA; T NANAYAMA; T TANABE

    1995年, PROSTAGLANDINS AND RELATED COMPOUNDS, 23, 109 - 111, web_of_science

    研究論文(学術雑誌)

  • STRUCTURE AND FUNCTION OF RENIN BINDING-PROTEIN

    S TAKAHASHI; H INOUE; K FUKUI; Y MIYAKE

    1994年12月, KIDNEY INTERNATIONAL, 46 (6), 1525 - 1527, web_of_science

    研究論文(学術雑誌)

  • THE CYCLIC-AMP RESPONSE ELEMENT PLAYS AN ESSENTIAL ROLE IN THE EXPRESSION OF THE HUMAN PROSTAGLANDIN ENDOPEROXIDE SYNTHASE-2 GENE IN DIFFERENTIATED U937 MONOCYTIC CELLS

    H INOUE; T NANAYAMA; S HARA; C YOKOYAMA; T TANABE

    The promoter activity of 1432 bp upstream of the human prostaglandin-endoperoxide synthase 2 gene (PTGS2) was examined in differentiated U937 monocytic cells expressing prostaglandin-endoperoxide synthase 2 mRNA. Transient transfection experiments were performed using these cells and reporter vectors containing the upstream region of the gene with deletions or site-specific mutations and the luciferase gene. The deletion or destruction of the cyclic AMP response element (nucleotides -59 to -53) markedly reduced the promoter activity of this gene. Electrophoretic mobility shift assays showed that a nuclear protein(s) binding to the cyclic AMP response element was induced during monocytic differentiation of U937 cells. These results indicate that expression of the human prostaglandin-endoperoxide synthase 2 gene in differentiated U937 monocytic cells is regulated by the cyclic AMP response element., 1994年08月, FEBS LETTERS, 350 (1), 51 - 54, doi;web_of_science

    研究論文(学術雑誌)

  • ISOLATION AND MOLECULAR-CLONING OF PROSTACYCLIN SYNTHASE FROM BOVINE ENDOTHELIAL-CELLS

    S HARA; A MIYATA; C YOKOYAMA; H INOUE; R BRUGGER; F LOTTSPEICH; ULLRICH, V; T TANABE

    Prostacyclin synthase catalyzes the conversion of prostaglandin H-2 to prostacyclin, which is a powerful vasodilator and the most potent natural occurring in hibitor of platelet aggregation. In the present study, we determined the amino acid sequence of bovine prostacyclin synthase by combined protein chemical and molecular cloning techniques. The enzyme was purified and characterized from bovine aorta microsomes, and the partial amino acid sequences were determined with the native enzyme and endoproteinase Lys-C-cleaved peptides. Using primers synthesized according to the amino acid sequences, cDNA coding for prostacyclin synthase was amplified by polymerase chain reaction with bovine endothelial cell poly(A)(+) RNA and cloned into pBluescript II. Nucleotide sequence analyses of the cloned cDNA inserts revealed that cDNA for this enzyme contained a 1500-base pair open reading frame coding for a 500-amino acid polypeptide with a M(r) of 56,628. COS-7 cells transfected with an expression plasmid harboring this cDNA clone expressed prostacyclin synthase activity. The primary structure of the enzyme showed structural characteristics of cytochrome P450 and exhibited a 32% identity to that of human cholesterol 7 alpha-hydroxylase. However, the identity between the amino acid sequences of bovine prostacyclin synthase and human thromboxane synthase was only 16%, and no P450 showed an identity higher than 40%, suggesting that prostacyclin synthase represents a new family in the P450 superfamily. RNA blot analysis indicated that the mRNA for prostacyclin synthase from bovine endothe lial cells showed a size of approximately 2.7 kilobases and that the mRNA level increased about 3-fold by treat ment of tumor necrosis factor-alpha., 1994年08月, JOURNAL OF BIOLOGICAL CHEMISTRY, 269 (31), 19897 - 19903, web_of_science

    研究論文(学術雑誌)

  • MOLECULAR-CLONING AND EXPRESSION OF HUMAN PROSTACYCLIN SYNTHASE

    A MIYATA; S HARA; C YOKOYAMA; H INOUE; ULLRICH, V; T TANABE

    The cDNA for human prostacyclin synthase was cloned by polymerase chain reaction using poly(A)(+) RNA from human aortic endothelial cells according to the partial nucleotide sequence of prostacyclin synthase gene. The cloned cDNA with a size of 1977 base pairs contained a 1500 base pairs open reading frame which encoded a 500 amino acid protein sharing an 88 % identity with bovine prostacyclin synthase. RNA blot analysis indicated that the size of major prostacyclin synthase mRNA of human aortic endothelial cells was approximately 6 kilobases and that its mRNA level was increased by interleukin 1 or interleukin 6 treatment. Moreover, tissue distribution study demonstrated that prostacyclin synthase mRNA is widely expressed in human tissues and is particularly abundant in ovary, heart, skeletal muscle, lung, and prostate. These results suggest a variety of physiological roles of prostacyclin in addition to the implications in the cardiovascular system. (C) 1994 Academic Press, Inc., 1994年05月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 200 (3), 1728 - 1734, doi;web_of_science

    研究論文(学術雑誌)

  • ENHANCER CONTAINING UNUSUAL GC BOX-LIKE SEQUENCES ON THE HUMAN CHOLINE-ACETYLTRANSFERASE GENE

    H INOUE; EE BAETGE; LB HERSH

    The minimum requirement of an enhancer for the human choline acetyltransferase (ChAT) gene was analyzed by mutagenesis and protein-DNA interaction studies. A series of deletion and site-specific mutants were expressed transiently in a rat cholinergic neuroblastoma cell, NS20Y. The results revealed that the distal region between -970 and -941 base pairs (bp) from the transcription start site is essential for efficient transcription of the human ChAT gene. Two GC box-like sequences were located within this region, and they were shown to bind purified Sp1 transcription factor by footprinting and gel retardation analyses. This sequence, however, has an orientational effect and is located approximately 1000 bp from the transcription start site, unusual for the conventional GC box sequence. Furthermore, the sequence between these GC box-like sequences is shown to bind another novel trans-acting factor by gel retardation assay and to be necessary for efficient enhancer activity. On the other hand, gel retardation assays using a nuclear extract from Drosophila SL2 cells suggested that non-Spl trans-acting factors could also participate in the enhancing activity of this element. Taken together, the results demonstrate that two GC box-like sequences and another trans-acting factor-binding sequence located between -970 and -941 bp act coordinately and are essential for efficient transcription of this low expressed human ChAT gene., 1993年12月, MOLECULAR BRAIN RESEARCH, 20 (4), 299 - 304, web_of_science

    研究論文(学術雑誌)

  • THE HUMAN GENE FOR RENIN-BINDING PROTEIN

    S TAKAHASHI; H INOUE; Y MIYAKE

    The human renin-binding protein (RnBP) gene was isolated from a human placental genomic library and characterized. The gene spans about 10 kilobases and consists of 11 exons separated by 10 introns. The 5'-flanking region and the exon-intron boundaries were sequenced. Residue G* in 5'-CGAG*TGG-3' was identified as the major transcription initiation site, and "GC" boxes were found in the vicinity of the cap site. No typical "TATA" or "CCAAT" box exists in the 5'-flanking region. The hydrophobic domain followed by a leucine-zipper motif in RnBP is encoded by the sixth exon. A fragment of the human RnBP gene (nucleotides -739 to +244) linked to the chloramphenicol acetyltransferase gene was transfected into human Wilms' tumor G401 and mouse L929 cells. The expression of this chimeric gene in G401 cells was 4-fold higher than that in L929 cells, the tissue-specific regulation of RnBP gene expression thus being suggested. The promoter for the RnBP gene was shown to be localized in nucleotides -35 to +244 on assaying of the promoter activity using deletion mutants of the chimeric constructs. The RnBP gene was found to be located in human chromosome X by means of polymerase chain reaction of hybrid DNAs from human and hamster somatic cells., 1992年06月, JOURNAL OF BIOLOGICAL CHEMISTRY, 267 (18), 13007 - 13013, web_of_science

    研究論文(学術雑誌)

  • MODULATION OF ACTIVE RENIN SECRETION BY RENIN-BINDING PROTEIN (RNBP) IN MOUSE PITUITARY ATT-20 CELLS TRANSFECTED WITH HUMAN RENIN AND RNBP CDNAS

    H INOUE; S TAKAHASHI; Y MIYAKE

    To investigate the role of renin-binding protein (RnBP) in renin metabolism, RnBP expression plasmid, which was constructed to express human RnBP under the control of mouse mammary tumor virus long terminal repeat, was transfected into mouse pituitary AtT-20 cells together with the expression plasmid encoding human renin. The transfectant secreted prorenin and active renin, whereas RnBP was expressed only in the presence of dexamethasone and without secretion into the medium. The secretion of active renin was stimulated by forskolin, and the stimulation was repressed by dexamethasone. The secretion of prorenin, however, was insensitive to forskolin irrespective of the presence or absence of dexamethasone. Moreover, the forskolin-stimulated release of active renin was hardly repressed by dexamethasone in AtT-20 cells transfected with the renin expression plasmid and a selectable plasmid pMAMneo. Coexistence of RnBP and renin mRNAs in human Wilms' tumor G-401 cells was shown by means of polymerase chain reaction of respective cDNAs from the cells. These results suggest that RnBP modulates the release of active renin in renin-producing cells., 1992年03月, JOURNAL OF BIOCHEMISTRY, 111 (3), 407 - 412, web_of_science

    研究論文(学術雑誌)

  • GENETIC AND MOLECULAR-PROPERTIES OF HUMAN AND RAT RENIN BINDING-PROTEINS WITH REFERENCE TO THE FUNCTION OF THE LEUCINE ZIPPER MOTIF

    H INOUE; S TAKAHASHI; K FUKUI; Y MIYAKE

    The presence of a leucine zipper motif was recognized in the deduced amino acid sequences of human and rat renin-bindig proteins (RnBPs) on cloning and sequence analysis of the RnBP cDNAs. The in vitro synthesized RnBPs, with the respective cDNAs, formed heterodimers with porcine renin and homodimers. On comparison of these properties with those of porcine RnBP, the leucine zipper motif was suggested to be a functional domain common to animal RnBPs. In addition to the motif, a hydrophobic domain adjacent to the motif and 10 cysteine residues were also well conserved in the three RnBPs. Moreover, about 85% of their amino acid sequences were identical. The RnBP mRNAs were expressed in the kidneys as the same size of 1.5-kb and the genes are suggested to exist as single copies in the genomes. Despite the high similarities in genetic and molecular properties, the molecular weights of human and rat RnBPs were 43,000, which is 1,000 larger than that of porcine RnBP. The immunoreactivities of human and rat RnBPs toward anti-porcine RnBP antiserum were 88 and 8% that of porcine RnBP, respectively, and the affinities of the two RnBPs for porcine renin were remarkably less than that of porcine RnBP. Moreover, the human and rat RnBP homodimers were partly dissociated under the conditions under which porcine RnBP existed as a dimer. These results indicate distinct differences in the molecular properties among the three RnBPs, in spite of their being highly similar structurally and functionally., 1991年10月, JOURNAL OF BIOCHEMISTRY, 110 (4), 493 - 500, web_of_science

    研究論文(学術雑誌)

  • LEUCINE ZIPPER MOTIF IN PORCINE RENIN-BINDING PROTEIN (RNBP) AND ITS RELATIONSHIP TO THE FORMATION OF AN RNBP-RENIN HETERODIMER AND AN RNBP HOMODIMER

    H INOUE; S TAKAHASHI; K FUKUI; Y MIYAKE

    An apparent leucine zipper motif was recognized in the predicted amino acid sequence of porcine kidney renin-binding protein (RnBP) by analysis of the nucleotide sequence of a cDNA encoding the protein (Inoue, H., Fukui, K., Takahashi, S., and Miyake, Y. (1990) J. Biol. Chem. 265, 6556-6561). To evaluate the role of this motif in the formation of an RnBP-renin heterodimer and an RnBP homodimer, a porcine mutant cDNA involving Leu185 --> Asp and Leu192 --> Asp substitutions was constructed and expressed in vitro and in Xenopus oocytes. The mutant protein neither binds to renin nor forms the homodimer. The results strongly suggest that the leucine zipper motif in the RnBP molecule mediates the formation of both the RnBP-renin heterodimer and the RnBP homodimer observed previously. The existence of the motif should facilitate elucidation of the role of RnBP in renin metabolism., 1991年06月, JOURNAL OF BIOLOGICAL CHEMISTRY, 266 (18), 11896 - 11900, web_of_science

    研究論文(学術雑誌)

  • MOLECULAR-CLONING AND SEQUENCE-ANALYSIS OF A CDNA-ENCODING A PORCINE KIDNEY RENIN-BINDING PROTEIN

    H INOUE; K FUKUI; S TAKAHASHI; Y MIYAKE

    1990年04月, JOURNAL OF BIOLOGICAL CHEMISTRY, 265 (12), 6556 - 6561, web_of_science

    研究論文(学術雑誌)

  • BIOSYNTHESIS OF A RENIN BINDING-PROTEIN

    K FUKUI; H INOUE; S TAKAHASHI; Y MIYAKE

    1989年10月, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 164 (1), 265 - 270, doi;web_of_science

    研究論文(学術雑誌)

  • IDENTIFICATION AND STRUCTURE OF THE RAT TRUE TISSUE KALLIKREIN GENE EXPRESSED IN THE KIDNEY

    H INOUE; K FUKUI; Y MIYAKE

    1989年05月, JOURNAL OF BIOCHEMISTRY, 105 (5), 834 - 840, web_of_science

    研究論文(学術雑誌)

  • Rat pyruvate kinase M gene. Its complete structure and characterization of the 5'-flanking region

    M. Takenaka; T. Noguchi; H. Inoue; K. Yamada; T. Matsuda; T. Tanaka

    Genomic clones containing the rat pyruvate kinase M gene, which encodes the M1- and M2-type isozymes, were isolated and their exon sequences were determined. This gene contains 12 exons and 11 introns and is 20 kilobases (kb) long. The sequences specific to the M1- and M2-types exist in exons 9 and 10, respectively (Noguchi, T., Inoue, H., and Tanaka, T. (1986) J. Biol. Chem. 261, 13807-13812). The seventh intron begins with the GC dinucleotide instead of the consensus GT dinucleotide, but other exon-intron boundaries are consistent with the 'GT-AG' rule. S1 mapping analysis showed that M1- and M2-type mRNAs had multiple, but the same transcription initiation sites. Thus, the M1- and M2-type isozyme mRNAs are concluded to be produced from the same M gene transcript by alternative RNA splicing. RNA blot hybridization analysis indicated that developmental changes of the isozymes in brain and skeletal muscle were regulated at the level of RNA splicing. The 5'-flanking region of the gene has no 'TATA box' or 'CAAT box', but contains potential Sp1 binding sites. Bacterial chloramphenicol acetyltransferase assay revealed that a fragment of about 0.5 kb of the 5'-flanking region of the gene was sufficient for promoter activity in the rat hepatoma cell line, dRLh-84. This activity was not present in adult rat hepatocytes, indicating that the 0.5-kb fragment has tissue-specific promoter activity. A processed-type pseudogene that resembles the M2-type pyruvate kinase cDNA was also characterized., 1989年, Journal of Biological Chemistry, 264 (4), 2363 - 2367, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • The L- and R-type isozymes of rat pyruvate kinase are produced from a single gene by use of different promoters.

    T. Noguchi; K. Yamada; H. Inoue; T. Matsuda; T. Tanaka

    cDNA clones for rat R-type pyruvate kinase and a genomic clone encoding both L- and R-type isozyme mRNAs were isolated. Their sequences were compared with that of the L-type isozyme cDNA to determine the sequences of mRNA and protein of the R-type isozyme and the organizations of the L- and R-type genes. Results showed that the R-type isozyme mRNA had an identical nucleotide sequence to that of the L-type except in the 5'-terminal region including the coding sequence and the length of the 3'-untranslated region. The sequence upstream of the 5th coding residue of the L-type was replaced by a 98-nucleotide coding sequence plus a 5'-untranslated region in the R-type isozyme. Therefore, the R-type subunit consists of 574 amino acids, which is 31 residues longer than the L-type at the amino terminus. The pyruvate kinase L gene is present as a single copy per haploid genome and is composed of 12 exons and 11 introns with a length of about 9.3 kilobase pairs. The first (exon R) and second (exon L) exons encode the 5'-terminal sequences specific for the R- and L-types, respectively. The remaining downstream exons encode a sequence common to both isozymes. The last exon contains the entire 3'-untranslated region, including several putative polyadenylation signals. Alternative use of these signals is reported to be responsible for generation of multiple mRNA species for the L-type, whereas the R-type uses only the first signal. The cap site is mapped 16 nucleotides upstream from the translation initiation site for the L-type, whereas multiple cap sites were suggested for the R-type. The canonical promoter of the TATA box was identified in the upstream sequence of exon L, but not in that of exon R. Instead, the 5'-flanking region of exon R contained another promoter sequence of the CAT box. Thus, we conclude that the L- and R-type isozymes of pyruvate kinase are produced from a single gene by use of different promoters., 1987年10月15日, The Journal of biological chemistry, 262 (29), 14366 - 14371, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Complete amino acid sequence of rat L‐type pyruvate kinase deduced from the cDNA sequence

    Hiroyasu INOUE; Tamio NOGUCHI; Takehiko TANAKA

    cDNA clones, containing the entire coding region of rat L‐type pyruvate kinase, were isolated and their nucleotide sequences were determined by the dideoxy‐chain‐termination method. The predicted coding region, which spans 543 amino acids, established the complete amino acid sequence of the L‐type isozyme of pyruvate kinase for the first time. The deduced amino acid sequence of the L type has one phosphorylation site in its amino terminus and shows about 68% and 48% homologies with M1‐type pyruvate kinase of chicken and yeast pyruvate kinase respectively. Domain A exhibits higher homology than domains B and C. The residues in the active site of the L‐type enzyme of rats, lying between domains B and A2, are rather different from those of the M1‐type enzyme of chickens, but other residues constituting the active site are identical with those of the chicken M1 type except for one amino acid substitution. Copyright © 1986, Wiley Blackwell. All rights reserved, 1986年01月, European Journal of Biochemistry, 154 (2), 465 - 469, doi;pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • The M1- and M2-type isozymes of rat pyruvate kinase are produced from the same gene by alternative RNA splicing

    T. Noguchi; H. Inoue; T. Tanaka

    The complete nucleotide sequences of rat M1- and M2-type pyruvate kinase mRNAs were determined by sequencing the cDNAs and by analyses of S1 nuclease mapping and primer extension. The sequences have an identical molecular size of about 2220 nucleotides excluding a poly(A) tail and include 1593-nucleotide coding region. Their nucleotide sequences are identical except for 160-nucleotide sequences within the coding regions. The amino acid sequences of the M1- and M2-type subunits deduced from the cDNA sequences differ by only 45 residues within domain C, which constitutes the main region responsible for intersubunit contact. The sequence of this region of the M2-type shows higher homology than that of the M1-type with the corresponding sequence of the L-type. Since the M2- and L-types are allosteric enzymes, unlike to the M1-type, the residues common to the M2- and L-types, but not to the M1-type may be important for mediating the allosteric properties. Genomic clones encoding both M1- and M2-type isozyme mRNAs were isolated. By partial sequence analysis of a clone λMPK37 four exons were indentified, of which two adjacent exons coded the M1- and M2-specific sequences, respectively. The two remaining exons present downstream coded amino acids common to the two isozymes. Thus, we conclude that the M1- and M2-type isozymes of pyruvate kinase are produced from the same gene probably by alternative RNA splicing., 1986年, Journal of Biological Chemistry, 261 (29), 13807 - 13812, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Transcriptional and post-transcriptional regulation of L-type pyruvate kinase in diabetic rat liver by insulin and dietary fructose

    T. Noguchi; H. Inoue; T. Tanaka

    Regulation of the expression of the hepatic L-type pyruvate kinase gene by insulin and dietary fructose was studied in diabetic rats. Insulin increased the levels of putative nuclear RNA precursor species of this enzyme in parallel with that of total cellular pyruvate kinase L mRNA. These changes occurred more slowly than those induced by dietary fructose. Insulin caused a 3-fold increase in transcription of the pyruvate kinase L gene after 6 h and a 6-fold increase after 16 h. The increase caused by insulin was inhibited by glucagon, but not by adrenalectomy. Cycloheximide inhibited the induction caused by insulin, suggesting that insulin may stimulate transcription of the pyruvate kinase L gene by stimulating synthesis of some unknown protein. On the other hand, feeding fructose had no effect on transcription of the pyruvate kinase L gene. We previously showed that increases in the levels of putative nuclear RNA precursor species of the pyruvate kinase L after fructose feeding preceded changes in the levels of cytosolic pyruvate kinase L mRNA (Inoue, H., Noguchi, T., and Tanaka, T. (1984) J. Biochem. (Tokyo) 96, 1457-1462). Thus, dietary fructose may increase the levels of pyruvate kinase L mRNA by stabilizing nuclear RNA species. Glucagon inhibited the increase in pyruvate kinase L mRNA caused by dietary fructose. However, plasma levels of glucagon and thyroid hormones were not decreased in diabetic rats after fructose feeding. In addition, treatment with triiodo-L-thyronine caused no change in the pyruvate kinase L mRNA level. Furthermore, adrenalectomy did not impair enzyme induction by fructose in diabetic rats. Thus, the effect of fructose on pyruvate kinase L seems to be directly on the liver., 1985年, Journal of Biological Chemistry, 260 (26), 14393 - 14397, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Rapid regulation of L-type pyruvate kinase mRNA by fructose in diabetic rat liver

    Hiroyasu Inoue; Tamio Noguchi; Takehiko Tanaka

    The effect of fructose on the induction of L-type pyruvate kinase mRNA in diabetic rat liver was studied by using a cloned cDNA probe. Fructose feeding resulted in a 5- to 6-fold increase in the L-type enzyme mRNA level after 1 to 3 days. These changes were approximately proportional to the changes in the level of translatable mRNA of this enzyme. A significant increase in total cellular L-type enzyme mRNA level was observed within 2 h after fructose feeding and the level reached a maximum after 8 h. Dietary glycerol also markedly increased the L-type mRNA level. These alterations were essentially due to the changes in the cytosolic mRNA. Northern blot analysis of total cellular RNA revealed that two L-type enzyme mRNA species with molecular sizes of 2.1 and 3.6 kilobases were proportionally increased during the fructose induction. The two mRNA forms were found in immunopurified L-type enzyme mRNA and directed synthesis of the L-type subunit in vitro; they are therefore functional mature forms. In contrast, analysis of nuclear RNA showed five putative precursor RNA species for the enzyme, up to 9.4 kilobases in length, in the liver of fructose-fed rats, while no band of the RNA species was found in the nuclei of control liver. The changes in the number of bands of these RNA species and their intensities after fructose feeding preceded the changes in the level of total cellular L-type enzyme mRNA sequences. These results indicate that dietary fructose causes a rapid increase in the level of L-type pyruvate kinase mRNA sequences by acting at the nuclear level. © 1984, by the Japanese Biochemical Society., 1984年10月, Journal of Biochemistry, 96 (5), 1457 - 1462, doi;pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Molecular cloning of cDNA sequences for rat M2-type pyruvate kinase and regulation of its mRNA

    T. Noguchi; H. Inoue; Y. Nakamura; H. L. Chen; K. Matsubara; T. Tanaka

    Rat M2-type pyruvate kinase mRNA was enriched from total polysomes isolated from AH-130 Yoshida ascites hepatoma cells, which contain a very high concentation of the M2-type enzyme, by immunoprecipitation with a specific antibody and Staphylococcus aureus cells. Double-stranded cDNA synthesized from the enriched mRNA was inserted into the PstI site of pBR322, and the resultant recombinant DNA molecules were used to transform Escherichia coli. Three clones containing DNA complementary to M2-type pyruvate kinase mRNA were identified by colony hybridization, hybrid-arrested translation, and hybrid-selected translation. A partial restriction map was constructed covering about 1.44 kilobase pairs. The cloned region of the M2-type mRNA showed a high degree of sequence homology with the M1-type mRNA and some homology with the L-type mRNA as determined by dot blot hybridization. The molecular size of the M2-type mRNA, which was estimated to be 2.35-2.65 kilobases on denaturing gel, was the same as that of the M1-type mRNA. The level of hepatic M2-type pyruvate kinase mRNA measured by hybridization assay using cloned cDNA as a probe was increased 2.5-fold 1 day and 3.9-fold 2 days after partial hepatectomy and then started to decrease. This induction was followed by similar changes in the enzyme activity. AH-130 hepatoma cells contained 100-150 times more M2-type isoenzyme mRNA than regenerating liver. These results suggest that the increased levels of M2-type isoenzyme in regenerating liver and hepatoma cells are primarily due to elevation of hybridizable M2-type mRNA concentration., 1984年, Journal of Biological Chemistry, 259 (4), 2651 - 2655, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Molecular cloning of DNA complementary to rat L-type pyruvate kinase mRNA. Nutritional and hormonal regulation of L-type pyruvate kinase mRNA concentration

    T. Noguchi; H. Inoue; H. L. Chen; K. Matsubara; T. Tanaka

    Rat liver L-type pyruvate kinase mRNA was enriched from total polysomes by immunoprecipitation with a specific antibody and Staphylococcus aureus cells. Double-stranded cDNA synthesized from the enriched mRNA was inserted into the PstI site of pBR322, and the resultant recombinant DNA molecules were used to transform Escherichia coli. Three clones containing DNA complementary to L-type pyruvate kinase mRNA were identified by colony hybridization, hybrid-selected translation, and dot blot hybridization. A partial restriction endonuclease map of cDNA inserts was constructed covering about 1.86 kilobase pairs. The cDNA insert of recombinant plasmid pLPK-14 was used as a hybridization probe to quantitate L-type pyruvate kinase mRNA in rat liver after various treatments. The level of hybridizable L-type enzyme mRNA was markedly increased by a high carbohydrate diet. Diabetes greatly reduced the mRNA level in the liver of rats maintained on a high carbohydrate diet, but insulin administration resulted in restoration of the mRNA level to normal within 24 h. These changes were approximately proportional to the changes in the level of translatable L-type pyruvate kinase mRNA. Thus, we conclude that nutrition and hormonal regulation of synthesis of hepatic L-type pyruvate kinase occurs at the pretranslational level., 1983年, Journal of Biological Chemistry, 258 (24), 15220 - 15223, pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

  • Regulation of Rat Liver L‐Type Pyruvate Kinase mRNA by Insulin and by Fructose

    Tamio NOGUCHI; Hiroyasu INOUE; Takehiko TANAKA

    The effects have been studied of streptozotocin‐induced diabetes and subsequent insulin administration or feeding of a high fructose diet on the amount of enzyme protein and mRNA activity of L‐type pyruvate kinase in rat liver. Diabetes markedly decreased the L‐type enzyme activity in rat liver and insulin treatment resulted in restoration of the enzyme activity to normal. A high fructose diet also increased the enzyme activity in diabetic rats but to a lesser extent. Immunochemical analysis showed that these alterations in the enzyme activity were due to changes in the amount of immunoreactive enzyme protein. The mechanism of the changes was studied further by assaying the level of functional mRNA coding for this enzyme in a nuclease‐treated reticulocyte lysate system with total RNA isolated from rat liver. L‐type pyruvate kinase mRNA, expressed as a percentage of the total protein synthesized, was greatly decreased in diabetic rats. Insulin administration resulted in recovery of the mRNA activity to the normal level within 24 h. The lag period before accumulation of translatable mRNA of the L‐type enzyme was about 4‐5 h. The mRNA activity was also increased in diabetic rats fed a high fructose diet. This fructose effect, which was much smaller than the insulin effect, was maximal after feeding fructose diet for one day. These changes were approximately comparable to the changes in enzyme activity. Thus, it is suggested that regulations of rat liver L‐type pyruvate kinase by insulin and by fructose are primarily due to changes in the level of translatable mRNA of this enzyme. Copyright © 1982, Wiley Blackwell. All rights reserved, 1982年11月, European Journal of Biochemistry, 128 (2-3), 583 - 588, doi;pubmed;scopus;scopus_citedby

    研究論文(学術雑誌)

MISC

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  • 葉酸欠乏が脂質代謝関連酵素の遺伝子発現に及ぼす影響

    井上 裕康; 中田理恵子

    2008年, 脂質生化学研究, 50, 300 - 303

  • レスベラトロール四量体バチカノールCによる核内受容体PPARα及びβ/δの活性化

    井上 裕康; 塚本朋子他

    2008年, 脂質生化学研究, 50, 304 - 307

  • PPARと誘導型シクロオキシゲナーゼの関係

    井上 裕康

    2007年, 医学のあゆみ, 220 (1), 93 - 98

  • 植物性ポリフェノールにおける核内受容体PPARの活性化

    中田 理恵子; 井上裕康; 中田理恵子

    2006年, 脂質生化学研究, 48, 141 - 143

  • ナノレベルイメージングによる分子構造と機能の解析

    井上 裕康; 盛 英三; 望月 直樹; 武田 壮一; 井上 裕康; 中村 俊; 土屋 利江

    2006年, 日本臨床, 64, 358 - 364

  • 疾患関連蛋白のサブナノ構造イメージングと分子標的薬剤の開発; ナノイメージング構造

    井上 裕康; 盛 英三; 武田 壮一; 若林 繁夫; 井上 裕康; ユーセフベンアマー; 松原孝宣; 五十嵐智子; 柴田 洋之

    2006年, 分子心血管病, 7, 340 - 346

  • 植物性ポリフェノールによる核内受容体PPARの活性化

    井上 裕康; 井上 裕康; 中田理恵子; 竹内悠; 塚本朋子; 堀田真理子; 名村尚武

    2006年, 脂質生化学研究, 48, 131 - 134

  • 疾患関連蛋白のサブナノ構造イメージングと分子標的薬剤の開発; ナノイメージング構造

    INOUE HIROYASU; 盛 英三; 武田 壮一; 若林 繁夫; 井上 裕康; ユーセフベンアマー; 松原孝宣; 五十嵐智子; 柴田 洋之

    2006年, 7, 340 - 346

  • T-oligo decreases COX-2 expression via p53 and NF kappa B-dependent promoter repression

    Marwaha, V; S Arad; B Helms; H Inoue; R Der Sarkissian; BA Gilchrest; DA Goukassian

    BLACKWELL PUBLISHING INC, 2005年04月, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 124 (4), A90 - A90, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • PPARsの内因性リガンド

    井上 裕康; 井上 裕康

    2005年, 日本臨床, 63 (4), 578 - 583

  • 「フレンチパラドックス」と核内受容体PPARとの新しい接点

    井上 裕康; 井上 裕康

    2005年, 化学と生物, 43, 619 - 624, doi

  • Gene expression induced by spreading depression is suppressed by selective COX-2 inhibitor JTE-522

    C Yokota; Y Kuge; H Inoue; N Tamaki; K Minematsu

    LIPPINCOTT WILLIAMS & WILKINS, 2004年06月, STROKE, 35 (6), E253 - E253, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • T-Oligo treatment down-regulates constitutive and UV-induced COX-2 expression in cultured cells and in murine and human skin

    DA Goukassian; G Marwaha; S Arad; B Helms; H Inoue; R Der Sarkissian; BA Gilchrest

    BLACKWELL PUBLISHING INC, 2004年03月, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 122 (3), A27 - A27, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • 赤ワインに含まれるポリフェノール・レスベラトロールに関する最近の話題

    井上 裕康; 井上 裕康

    2004年, ビタミン, 78, 621 - 623, doi

  • Characterization of I-123-Iomazenil distribution in a rat model of focal cerebral ischemia in comparison with DNA fragmentation.

    T Kaji; Y Kuge; C Yokota; M Tagaya; H Inoue; T Shiga; K Minematsu; N Tamaki

    SOC NUCLEAR MEDICINE INC, 2003年05月, JOURNAL OF NUCLEAR MEDICINE, 44 (5), 217P - 218P, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • ConstitutiveIy activated MEK1 represses sodium butyrate induced cell differentiation and apoptosis in rat intestinal epithelial cells

    K Komatsu; B Bose; JD Morrow; H Inoue; RN Dubois

    W B SAUNDERS CO, 2003年04月, GASTROENTEROLOGY, 124 (4), A598 - A598, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • Pretreatment with PPAR alpha agonists limits brain infarct after permanent focal cerebral ischemia in mice

    S Namura; K Umesono; XF Jiang; H Inoue

    LIPPINCOTT WILLIAMS & WILKINS, 2003年01月, STROKE, 34 (1), 299 - 299, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • Upregulation of cyclooxygenase-2 gene during spreading depression in a primate model

    C Yokota; H Inoue; Y Kuge; M Tagaya; Y Hasegawa; T Abumiya; N Ejima; N Tamaki; K Minematsu

    LIPPINCOTT WILLIAMS & WILKINS, 2003年01月, STROKE, 34 (1), 304 - 304, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • 核内受容体PPARを介する誘導型シクロオキシゲナーゼの発現調節に関する研究

    井上 裕康; Hiroyasu, Inoue

    2003年, ビタミン, 77, 449 - 458, doi

  • Characterization of I-123-iomazenil distribution in a rat model of focal cerebral ischemia - Comparison with COX-2 expression.

    T Kaji; Y Kuge; C Yokota; M Tagaya; H Inoue; T Shiga; K Minematsu; N Tamaki

    SOC NUCLEAR MEDICINE INC, 2002年05月, JOURNAL OF NUCLEAR MEDICINE, 43 (5), 237P - 238P, web_of_science

    研究発表ペーパー・要旨(国際会議)

  • PPARとプロスタグランジン生成

    井上 裕康; 井上 裕康

    2001年, 特集「PPARと疾患」 BIO Clinica, 16, 411 - 416

  • COX2の発現調節

    井上 裕康; 井上 裕康

    2001年, 医学のあゆみ 特集「COX2 基礎と臨床」, 197, 134 - 138

  • 第13章3節 遺伝子の構造とその発現調節

    井上 裕康; 井上 裕康; 田辺 忠

    2001年, 現代化学 増刊 プロスタグランジン研究の新展開 pp.122-127 (2001), 122 - 127

  • Transcriptional regulation of the human choline acetyltransferase gene

    LB Hersh; H Inoue; YP Li

    ELSEVIER SCIENCE BV, 1996年, CHOLINERGIC MECHANISMS: FROM MOLECULAR BIOLOGY TO CLINICAL SIGNIFICANCE, 109, 47 - 54, web_of_science

    書評論文,書評,文献紹介等

  • 誘導型シクロオキシゲナーゼ

    井上 裕康; 田辺 忠; 井上 裕康; 原 俊太郎

    1996年, ファルマシア, 32, 938 - 942

  • COX-1とCOX-2の生化学と分子生物学―誘導と転写調節機構

    井上 裕康; 井上 裕康; 田辺 忠

    1996年, 治療学, 30, 1347 - 1351

  • 新しい誘導型プロスタグランジン・エンドペルオキシド合成酵素 -遺伝子構造と発現調節機構を中心にして-

    井上 裕康; 井上 裕康; 横山 知永子; 田辺 忠

    1995年, 蛋白質核酸酵素, 40, 399 - 408

書籍等出版物

  • 分子栄養学

    井上裕康 (, 範囲: ヒトゲノム)

    講談社, 2018年

  • 非栄養素の分子栄養学

    中田理恵子; 井上裕康 (, 範囲: ポリフェノールによるPPAR機能制御と骨格筋代謝改善効果)

    建帛社, 2017年

  • 赤ワインの機能性成分レスベラトロールの分子標的と将来展望赤ワインの機能性成分レスベラトロールの分子標的と将来展望食品分析開発センターSUNATEC メールマガジン 2015年9月号 http://www.mac.or.jp/mail/150901/01.shtml

    井上裕康

    食品分析開発センターSUNATEC メールマガジン 2015年9月号, 2015年

  • 食品因子による栄養機能制御(日本栄養・食糧学会 監修)第14章 北海道で命名されたレスベラトロールのPPAR活性化を介した機能性

    井上裕康; 滝澤祥恵; 中田理恵子

    建帛社, 2015年 (ISBN: 9784767961811)

  • 分子栄養学 第8章 ヒトの遺伝子

    井上裕康

    羊土社, 2014年

  • 栄養・食糧学用語辞典改訂2版

    建帛社, 2014年

  • 食品学 I 改訂第2版 第6章 食品の機能性

    南江堂, 2011年

  • 食品学 Ⅱ 改訂第2版 第5章 油糧食品

    南江堂, 2011年

  • 食品学 II 改訂第2版 第8章 微生物利用食品

    中田理恵子; 井上裕康

    南江堂, 2011年

  • Techniques Used to Study Regulation of Cyclooxygenase-2 Promoter sites: Methods and Protocols, Methods in Molecular Biology , vol. 644, (2010)

    Springer, 2010年

  • Techniques Used to Study Regulation of Cyclooxygenase-2 Promoter sites: Methods and Protocols, Methods in Molecular Biology , vol. 644, (2010)

    Springer, 2010年

  • 食品学 I 第6章 食品の機能性

    南江堂, 2007年

  • 食品学 II 第5章 油糧食品

    南江堂, 2007年

  • 食品学 II 第8章 微生物利用食品

    南江堂, 2007年

  • General Food Science I (6) Functionality of Food

    Nankodo, 2007年

  • General Food Science II, (5) Food oil

    Nankodo, 2007年

  • General Food Science II, (8) Fermented Food

    Nankodo, 2007年

  • アラキドン酸カスケード Q&A (室田誠逸編)

    医薬ジャーナル社, 2002年

  • 第2章 COX遺伝子の解析

    『NOSとCOX:遺伝子から臨床へ』, 1996年

講演・口頭発表等

  • レスベラロール配糖体ピセイドによるPPARαを介した脂質代謝改善効果の検討

    第74回日本栄養・食糧学会大会, 2020年05月

    口頭発表(一般)

  • The direct activation of PPARalpha by 4’-hydroxyl group of resveratrol and a feedforward regulation via cAMP

    The 9th International Conference on Polyphenols and Health - ICPH2019, 2019年11月

  • レスベラトロールによるPPARα活性化とcAMPを介した制御

    第92回日本生化学会大会, 2019年09月

  • cAMPを介したレスベラトロールによるPPARα活性化の制御

    第61回日本脂質生化学会, 2019年07月

  • cAMPを介したレスベラトロールによるPPARα活性化

    第73回日本栄養食糧学会大会, 2019年05月

  • モリンガ種子抽出物の抗疲労効果の検証

    日本農芸化学会2019年度大会, 2019年03月

  • 運動とレスベラトロールによる骨格筋代謝改善効果

    第91回日本生化学会大会, 2018年09月

  • マウス由来初代培養心筋細胞に対するレスベラトロールの効果とPPARα活性化の関与

    第91回日本生化学会大会, 2018年09月

  • マウス由来初代培養心筋細胞に対するレスベラトロールの効果

    Food Congress 2018, 2018年09月

  • レスベラトロールと運動による骨格筋代謝改善効果

    Food Congress 2018, 2018年09月

  • マウス由来培養心筋細胞に対するレスベラトロールの効果

    第72回日本栄養・食糧学会大会, 2018年05月

  • Effect of resveratrol on human cardiomyocytes derived from iPS cells

    12th Asian Congress of Nutrition, 2015年

  • Possible participation of PPARalpha in improvement of lipid metabolism by soybean-derived components

    12th Asian Congress of Nutrition, 2015年

  • Effects on lipid metabolism by folate-deficiency in rats

    12th Asian Congress of Nutrition, 2015年

  • Screening of GPR120 activation by various long-chain fatty acids using TGFalpha shedding assay

    12th Asian Congress of Nutrition, 2015年

  • レスベラトロールによるヒトiPS細胞由来心筋細胞の遺伝子発現への影響

    日本家政学会 第67回大会, 2015年

  • PPAR活性化を指標とした大豆由来成分の脂質代謝改善効果の検討

    日本家政学会 第67回大会, 2015年

  • レスベラトロールによるPPARα活性化のcAMPを介するフィードフォワード制御

    第57回日本脂質生化学会大会, 2015年

  • TGFα切断アッセイを用いたω-3系脂肪酸によるGPR120活性化

    第67回日本ビタミン学会大会, 2015年

  • 母ラットの葉酸欠乏が出生仔に及ぼす影響

    第67回日本ビタミン学会大会, 2015年

  • cAMPを介したレスベラトロールによるPPARα活性化のfeedforward制御

    第9回日本ポリフェノール学会学術大会, 2015年

  • Feedforward activation of PPARα by resveratrol and its PDE inhibitory activity

    56th International Conference on the Bioscience of lipids, 2015年

  • 種々の脂肪酸によるGPR120活性化の検討

    第54回 日本栄養·食糧学会 近畿支部大会, 2015年

  • COX-2発現抑制とPPAR活性化を指標にした辛味成分の機能性評価

    第54回 日本栄養·食糧学会 近畿支部大会, 2015年

  • レスベラトロール長期摂取による抗動脈硬化作用の検討

    BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会), 2015年

  • マウスマクロファージ様RAW264細胞の極性化におけるPPARとCOX経路の関与の検討

    BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会), 2015年

  • GPR120活性化を指標とする新規食品機能評価系の検討

    BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会), 2015年

  • レスベラトロールによるPPARα活性化におけるcAMPを介したフィードフォワード制御

    BMB2015(第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会), 2015年

  • Effect of resveratrol on human cardiomyocytes derived from iPS cells

    12th Asian Congress of Nutrition, 2015年

  • Possible participation of PPARalpha in improvement of lipid metabolism by soybean-derived components

    12th Asian Congress of Nutrition, 2015年

  • Effects on lipid metabolism by folate-deficiency in rats

    12th Asian Congress of Nutrition, 2015年

  • Screening of GPR120 activation by various long-chain fatty acids using TGFalpha shedding assay

    12th Asian Congress of Nutrition, 2015年

  • Feedforward activation of PPARα by resveratrol and its PDE inhibitory activity

    56th International Conference on the Bioscience of lipids, 2015年

  • 葉酸欠乏による脂質代謝変動とコリンの影響

    第66回日本家政学会大会, 2014年

  • DNAチップ解析を用いたレスベラトロールによるヒト血管内皮細胞の遺伝子発現検討

    第66回日本家政学会大会, 2014年

  • 北海道で命名されたレスベラトロールのPPAR活性化を介した機能性

    第68回日本栄養・食糧学会大会(シンポジウム), 2014年

  • 培養心筋細胞を用いたレスベラトロールによる脂質代謝関連遺伝子発現の検討

    第68回日本栄養・食糧学会大会, 2014年

  • 培養細胞と生体における辛味成分によるPPARα活性化

    第68回日本栄養・食糧学会大会, 2014年

  • 大豆由来成分による脂質代謝改善効果とPPARα活性化

    第56回日本脂質生化学会大会, 2014年

  • 葉酸・コリン欠乏による肝脂肪蓄積の誘導

    第66回日本ビタミン学会大会, 2014年

  • PPAR活性化を介した辛味成分の機能性評価

    第66回日本ビタミン学会大会, 2014年

  • Induction of eNOS, SIRT1 and autophagy-related mRNAs in HUVEC by resveratrol

    International Conference of Polyphenol (ICP2014), 2014年

  • Resveratrol and its tetramer activate nuclear receptor PPARs in vitro and in vivo

    International Conference of Polyphenol (ICP2014), 2014年

  • HUVECにおけるレスベラトロールによるeNOS,SIRT1, オートファジー関連遺伝子の発現誘導

    日本生化学会, 2014年

  • PPARβ(δ) 遺伝子欠損マウスの行動表現型に関する検討

    第87回日本生化学会大会, 2014年

  • Resveratrol: a triple agonist for PPAR & a cell-type selective suppressor for COX-2 expression (Lecture 30 min)

    3rd International conference on resveratrol and health, 2014年

  • Induction of eNOS, SIRT1 and autophagy-related mRNAs in HUVEC by resveratrol

    International Conference of Polyphenol (ICP2014), 2014年

  • Resveratrol and its tetramer activate nuclear receptor PPARs in vitro and in vivo

    International Conference of Polyphenol (ICP2014), 2014年

  • Resveratrol: a triple agonist for PPAR & a cell-type selective suppressor for COX-2 expression (Lecture 30 min)

    3rd International conference on resveratrol and health, 2014年

  • 葉酸、コリン欠乏時における脂質代謝の変動

    第65回日本ビタミン学会大会, 2013年

  • COX-2発現抑制とPPAR活性化を介したシナモンバーク油の機能性評価

    第65回日本ビタミン学会大会, 2013年

  • シナモンバーク油による脂質代謝改善効果

    第65回日本家政学会大会, 2013年

  • 高脂肪食摂取時の葉酸欠乏が脂質代謝に及ぼす影響

    第65回日本家政学会大会, 2013年

  • シナモンバーク油によるPPARα活性化

    第67回日本栄養・食糧学会大会, 2013年

  • 酸味受容体候補PKD1L3/ PKD2L1に対するミラクリンの影響

    第67回日本栄養・食糧学会大会, 2013年

  • 低濃度レスベラトロールによるHUVECでの遺伝子発現変動

    第55回日本脂質生化学会, 2013年

  • 血管内皮細胞での低濃度レスベラトロール連続処理による eNOS, SIRT1, オートファジー関連遺伝子の発現誘導 2013年8月5日 第7回日本ポリフェノール学会年次大会 東京農工大学

    第7回日本ポリフェノール学会年次大会, 2013年

  • マウス系統差によるレスベラトロールPPARα活性化の相違

    第86回日本生化学会大会, 2013年

  • Evaluation of essential oil-derived components for prevention of lifestyle-related diseases targeted to COX-2 and PPAR

    ICN (International Congress of Nutrition) 2013, 2013年

  • Improvement of lipid metabolism and life span by resveratrol via PPARα activation

    ICN (International Congress of Nutrition) 2013, 2013年

  • Subcellualr localization of miraculin and its functional expression in E.coli

    ICN (International Congress of Nutrition) 2013, 2013年

  • Molecular mechanism of PPARα activation by resveratrol

    ICN (International Congress of Nutrition) 2013, 2013年

  • 葉酸欠乏によって誘導される脂質代謝の変動

    第52回日本栄養・食糧学会近畿支部大会, 2013年

  • レスベラトロールと運動による組織選択的PPARα活性化

    第341回脂溶性ビタミン総合研究委員会, 2013年

  • Evaluation of essential oil-derived components for prevention of lifestyle-related diseases targeted to COX-2 and PPAR

    ICN (International Congress of Nutrition) 2013, 2013年

  • Improvement of lipid metabolism and life span by resveratrol via PPARα activation

    ICN (International Congress of Nutrition) 2013, 2013年

  • Subcellualr localization of miraculin and its functional expression in E.coli

    ICN (International Congress of Nutrition) 2013, 2013年

  • Molecular mechanism of PPARα activation by resveratrol

    ICN (International Congress of Nutrition) 2013, 2013年

  • COX-2発現抑制、PPAR活性化を指標にしたニンニク油の機能性評価

    第64回日本家政学会大会, 2012年

  • 葉酸代謝酵素γ-グルタミルヒドロラーゼの臓器局在の検討

    第64回日本家政学会大会, 2012年

  • COX-2およびPPARを標的としたニンニク油の機能性評価

    第66回日本栄養・食糧学会大会, 2012年

  • PPARαを介したレスベラトロールの作用と系統差による効果の相違

    第66回日本栄養・食糧学会大会, 2012年

  • COX-2およびPPARを標的とした精油成分の機能性評価

    第54回日本脂質生化学会, 2012年

  • PPAR活性化を標的とするレスベラトロールの機能性評価と新しい食品素材の探索(基調講演)

    岡山バイオアクティブ研究会シンポジウム, 2012年

  • 葉酸欠乏における脂質代謝の変化—高脂肪付加の影響—

    第64回日本ビタミン学会大会, 2012年

  • 運動負荷とレスベラトロールによる組織選択的PPARα活性化

    第64回日本ビタミン学会大会, 2012年

  • レスベラトロール摂取と習慣的運動による筋肉PPARα活性化と持久力向上

    日本ポリフェノール学会 お茶の水女子大, 2012年

  • PPAR活性化を介するレスベラトロールの機能性

    第33回日本肥満学会 シンポジウム「食品の機能と肥満症」, 2012年

  • 習慣的運動とレスベラトロールによる筋肉PPARα活性化

    第33回日本肥満学会, 2012年

  • Cellular localization of Miraculin-YFP fusion protein

    第46回日本味と匂学会, 2012年

  • ミラクリンの分子進化に関する考察:細胞内局在の視点から

    日本栄養食糧学会近畿支部大会, 2012年

  • PPARα活性化によるレスベラトロールの脂質代謝改善効果

    第337回脂溶性ビタミン総合研究委員会, 2012年

  • シナモンバーク油による生体におけるPPARα活性化の検討

    第84回日本生化学会大会, 2012年

  • 味覚修飾タンパク質ミラクリンの細胞内局在におけるpHの影響

    第84回日本生化学会大会, 2012年

  • 葉酸とコリンの欠乏が高脂肪摂取時の脂質代謝に及ぼす影響

    第84回日本生化学会大会, 2012年

  • Cellular localization of Miraculin-YFP fusion protein

    2012年

  • レスベラトロール4’位水酸基はPPAR活性化に関与する

    第53回日本脂質生化学会 (ホテル東京ガーデンパレス), 2011年

  • PPARαを介したレスベラトロールの効果と系統差による脂質代謝の影響

    第53回日本脂質生化学会 (ホテル東京ガーデンパレス), 2011年

  • 細胞外分泌シグナル配列を持つ味覚修飾タンパク質ミラクリン

    第65回日本栄養・食糧学会大会 (お茶の水女子大), 2011年

  • COX-2およびPPARを標的としたシナモンバーク油の機能性評価

    第65回日本栄養・食糧学会大会 (お茶の水女子大), 2011年

  • レスベラトロールによるPPAR活性化における4’位水酸基の関与

    第65回日本栄養・食糧学会大会 (お茶の水女子大), 2011年

  • PPARαを介したレスベラトロールの作用機構と系統差による相違

    第63回日本家政学会大会 (和洋女子大), 2011年

  • レスベラトロール摂取と習慣的運動の相乗効果

    第63回日本家政学会大会 (和洋女子大), 2011年

  • 大腸菌によるγ‐グルタミルヒドロラーゼの発現とラット臓器での局在

    第62回日本ビタミン学会大会 (安田女子大学), 2011年

  • PPARαを介したレスベラトロールの作用機構 ‐系統差による相違の検討‐

    第62回日本ビタミン学会大会 (安田女子大学), 2011年

  • A possible novel function of Asian herbs for prevention of life-style related diseases targeted to COX-2 and PPARs

    ACN2011, 2011年

  • Resveratrol and its tetramer activate PPARs in vitro and in vivo

    ACN2011, 2011年

  • A possible function of resveratrol and essential oil-derived chemicals for prevention of life-style related diseases targeted to COX-2 and PPAR

    ICBL2011, 2011年

  • COX-2 - PPAR - Resveratrol

    第83回日本生化学会大会 シンポジウム:核内受容体と代謝, 2011年

  • 味覚修飾タンパク質ミラクリンの細胞内局在検討

    第83回日本生化学会大会, 2011年

  • レスベラトロール類似体によるPPARa活性化

    第83回日本生化学会大会, 2011年

  • 組換えγ‐グルタミルヒドロラーゼの発現と臓器局在

    第83回日本生化学会大会, 2011年

  • 運動負荷による筋肉でのレスベラトロールPPARα活性化

    第83回日本生化学会大会, 2011年

  • PPARとCOX-2を指標にした香辛料成分の機能評価

    第83回日本生化学会大会, 2011年

  • ミラクリンの酸味受容体PKD1L3/PKD2L1に対する影響の検討

    日本味と匂学会第45回大会(石川県立音楽堂), 2011年

  • COX-2とPPARを指標としたレスベラトロールの機能性評価 (シンポジウム)

    第5回 日本ポリフェノール学会年次大会 シンポジウム 「ポリフェノール研究の最前線」(座長:井上裕康、佐藤隆一郎) 木)東京大学弥生講堂, 2011年

  • A possible function of essential oil-derived chemicals for prevention of life-style related diseases targeted to COX-2 and PPAR. (poster award)

    ICOFF2011, 2011年

  • Activation of PPARs by resveratrol in vitro and in vivo.

    ICOFF2011, 2011年

  • ファイトケミカルとアンチエージング:薬食同源の視点から

    第34回日本分子生物学会ワークショップ「ファイトケミカルと疾病:その多彩な薬理学的作用」, 2011年

  • A possible novel function of Asian herbs for prevention of life-style related diseases targeted to COX-2 and PPARs

    ACN2011, 2011年

  • Resveratrol and its tetramer activate PPARs in vitro and in vivo

    ACN2011, 2011年

  • A possible function of resveratrol and essential oil-derived chemicals for prevention of life-style related diseases targeted to COX-2 and PPAR

    ICBL2011, 2011年

  • A possible function of essential oil-derived chemicals for prevention of life-style related diseases targeted to COX-2 and PPAR. (poster award)

    ICOFF2011, 2011年

  • Activation of PPARs by resveratrol in vitro and in vivo.

    ICOFF2011, 2011年

  • Phytochemicals and anti-aging: From a view of Drug-Food-Identical-Origin

    Phytochemicals and diseases: a wide variety of pharmacological functions, 2011年

  • 核内受容体PPARの活性化を指標とした食品機能成分のスクリーニングと運動の併用効果

    第14回 食と運動の機能性に関する研究会, 2010年

  • COX-2発現抑制とPPAR活性化を指標にした香辛料の機能性評価

    スパイス&ハーブ研究成果セミナー, 2010年

  • PPAR活性化とCOX-2発現抑制を指標にした食品機能成分の探索

    レドックス生命科学第170委員会第22回研究会, 2010年

  • PPAR活性化を指標にした食品機能成分の探索

    日本栄養・食糧学会シンポジウム 脂質・エネルギー代謝制御の分子栄養学, 2010年

  • PPAR活性化を指標にした食品機能成分の探索

    第64回 日本栄養・食糧学会大会 シンポジウム3: 脂質・エネルギー代謝制御の分子栄養学 オーガナイザー 井上 裕康 (奈良女子大学)、宮澤 陽夫 (東北大学)、佐藤隆一郎 (東京大学), 2010年

  • COX-2およびPPARを標的としたレモングラス油の機能性評価

    日本栄養・食糧学会, 2010年

  • in vivoにおけるPPARαを介したレスベラトロールの作用機構

    日本栄養・食糧学会, 2010年

  • レスベラトロール摂取が運動持久力に及ぼす影響

    日本栄養・食糧学会, 2010年

  • 大腸菌による組換えγ-グルタミルヒドロラーゼの発現と精製

    日本栄養・食糧学会, 2010年

  • COX-2発現抑制、PPAR活性化を指標にしたレモングラス精油の機能性評価

    第64回日本家政学会大会 広島大学, 2010年

  • Evaluation of Essential oils by activation of PPARs and suppression of COX-2 expression

    Keystone Symposium (Bioactive Lipids: Biochemistry and Diseases), 2010年

  • Resveratrol activates PPARs in vitro and in vivo

    Keystone Symposium (Bioactive Lipids: Biochemistry and Diseases), 2010年

  • PPARおよびCOX-2への作用を指標としたレモングラス精油成分の機能性評価

    第62回日本ビタミン学会大会(口頭) アイーナ(いわて県民情報交流センター), 2010年

  • 大腸菌による組換えγ‐グルタミルヒドロラーゼの発現と精製

    第62回日本ビタミン学会大会(口頭) アイーナ(いわて県民情報交流センター), 2010年

  • PPARαを介したレスベラトロールの作用機構 ‐個体レベルでの検討‐

    第52回日本脂質生化学会 森秋旅館(群馬県伊香保温泉), 2010年

  • PPARとCOX-2を標的とする抗酸化食品因子の機能探索

    第63回日本酸化ストレス学会 シンポジウム:抗酸化食品因子の新規な機能探索とその応用, 2010年

  • 核内受容体PPAR活性化を指標とした食品成分の探索と生体内での機能評価

    日本農芸化学会関西支部第465回講演会 ミニシンポジウム:食品成分と健康:生体応答の分子メカニズム, 2010年

  • COX-2とPPARを標的とした食品機能成分の機能探索

    第328回脂溶性ビタミン総合研究委員会, 2010年

  • 特殊なPPAR活性化能を示す香辛料シナモンバーク精油

    日本味と匂学会第44回大会 北九州国際会議場, 2010年

  • レスベラトロール四量体バチカノールCによるPPARα, β/δ活性化 -培養細胞および個体レベルでの検討-

    日本生化学会, 2010年

  • Evaluation of Essential oils by activation of PPARs and suppression of COX-2 expression

    Keystone Symposium (Bioactive Lipids: Biochemistry and Diseases), 2010年

  • Resveratrol activates PPARs in vitro and in vivo

    Keystone Symposium (Bioactive Lipids: Biochemistry and Diseases), 2010年

  • レスベラトロールがヒト臍帯静脈血管内皮細胞の遺伝子発現に及ぼす影響

    第63回日本栄養食糧学会, 2009年

  • 田村恵美、小菅由希子、刈谷斐、勝川路子、中田理恵子、井上裕康

    第63回日本栄養食糧学会, 2009年

  • レスベラトロール摂取による運動持久力改善効果の検討

    第63回日本栄養食糧学会, 2009年

  • 葉酸欠乏による肝臓及び脂肪組織での脂質代謝関連酵素の発現変動

    第63回日本栄養食糧学会, 2009年

  • Evaluation of essential oils by activation of PPARs and suppression of COX-2 expression

    5th Takeda Science Foundation Symposium on Pharma Sciences, 2009年

  • Evaluation of essential oils by activation of PPARs and suppression of COX-2 expression

    PLM2009, 2009年

  • PPARa及びCOX-2 への作用を指標としたバラ油成分の機能性評価

    第61回日本ビタミン学会, 2009年

  • 葉酸欠乏が肝臓及び脂肪組織における脂質代謝関連酵素の発現に及ぼす影響

    第61回日本ビタミン学会, 2009年

  • レスベラトロール摂取によるPPARαを介した作用機構

    第61回日本家政学会, 2009年

  • COX-2発現抑制、PPAR活性化を指標にした精油の機能性評価

    第61回日本家政学会, 2009年

  • シロイヌナズナにおけるミラクリン-YFP融合タンパク質の発現系構築

    第43回日本味と匂い学会, 2009年

  • COX-2発現抑制とPPAR活性化を指標にした香辛料成分の機能性評価

    第43回日本味と匂い学会, 2009年

  • Resveratrol activates nuclear receptor PPARs in vitro and in vivo (distinguished poster presentation)

    19th International Congress of Nutrition 2009, 2009年

  • レスベラトロール研究の歴史的背景

    日本生化学会 シンポジウム「レスベラトロールに関する最新の知見」 オーガナイザー井上裕康(奈良女子大)、堀尾嘉幸(札幌医大), 2009年

  • PPARを介したレスベラトロールの作用機構

    日本生化学会 シンポジウム「レスベラトロールに関する最新の知見」 オーガナイザー井上裕康(奈良女子大)、堀尾嘉幸(札幌医大), 2009年

  • PPAR活性化およびCOX-2発現抑制を指標としたバラ油の機能性評価

    日本生化学会, 2009年

  • in vivoにおけるレスベラトロールによるPPARα活性化

    日本生化学会, 2009年

  • レスベラトロール連続処理による血管内皮細胞の遺伝子発現変動

    日本生化学会, 2009年

  • レスベラトロールによるPPARαを介した運動持久力改善効果の検討

    日本生化学会, 2009年

  • 大腸菌を用いた組換えγ-グルタミルヒドロラーゼの発現と精製

    日本生化学会, 2009年

  • Carvacrol and other components of essential oils activate PPARs and suppress COX-2 expression

    Bioactive Lipid Conference, 2009年

  • Evaluation of essential oils by activation of PPARs and suppression of COX-2 expression

    5th Takeda Science Foundation Symposium on Pharma Sciences, 2009年

  • Evaluation of essential oils by activation of PPARs and suppression of COX-2 expression

    PLM2009, 2009年

  • Resveratrol activates nuclear receptor PPARs in vitro and in vivo (distinguished poster presentation)

    19th International Congress of Nutrition 2009, 2009年

  • Carvacrol and other components of essential oils activate PPARs and suppress COX-2 expression

    Bioactive Lipid Conference, 2009年

  • 核内受容体PPARαを介するブドウ新芽エキスによる運動持久力改善効果の可能性

    日本栄養食糧学会, 2008年

  • 葉酸欠乏による遺伝子発現の変化

    日本栄養食糧学会, 2008年

  • 味覚修飾タンパク質ミラクリンの発現とその機能評価

    日本生化学会近畿支部会, 2008年

  • ブドウ新芽エキス摂取による運動持久力改善効果の検討

    日本家政学会, 2008年

  • 遺伝子組換え技術による味覚修飾タンパク質ミラクリンの発現

    日本家政学会, 2008年

  • 葉酸欠乏が脂質代謝関連酵素の遺伝子発現に及ぼす影響

    日本脂質生化学会, 2008年

  • レスベラトロール四量体バチカノールCによる核内受容体PPARα及びβ/δの活性化

    日本脂質生化学会, 2008年

  • レスベラトロール四量体バチカノールCによる核内受容体PPARの活性化

    日本ビタミン学会, 2008年

  • 葉酸欠乏による遺伝子発現の変動

    日本ビタミン学会, 2008年

  • レスベラトロールとPPAR 現状と展望

    企業向け教育講演会, 2008年

  • PPARa activation by resveratrol and other polyphenols; Importance of 4-hydroxyl group

    The 22nd Naito Conference on Chemical Biology (I) Sapporo, Japan, 2008年

  • 大腸菌とシロイヌナズナを用いた組換えミラクリンの発現とその機能評価

    日本味と匂い学会, 2008年

  • 葉酸高含有卵(葉酸たまご)の生体内での有用性の評価

    日本家政学会関西支部, 2008年

  • COX-2およびPPARを標的とした精油成分の機能性評価

    日本栄養食糧学会近畿支部, 2008年

  • HUVECにおけるレスベラトロール連続処理による遺伝子発現変動の検討

    日本栄養食糧学会近畿支部, 2008年

  • 脳虚血障害後の環境刺激による神経機能回復に関する検討

    日本脳循環代謝学会総会, 2008年

  • 大腸菌を用いた味覚修飾活性を有する組換えミラクリンの発現

    日本生化学会, 2008年

  • レスベラトロール摂取によるPPARα依存的遺伝子群の誘導

    日本生化学会, 2008年

  • 葉酸欠乏における遺伝子発現の検討 ―肝臓および脂肪組織における脂質代謝関連酵素の変化―

    日本生化学会, 2008年

  • PPARa activation by resveratrol and other polyphenols; Importance of 4-hydroxyl group

    The 22nd Naito Conference on Chemical Biology (I) Sapporo, Japan, 2008年

  • ポリフェノールによるPPAR活性化と生活習慣病予防効果への検討

    2007年度日本農芸化学会大会, 2007年

  • レスベラトロール関連物質による核内受容体PPAR活性化と抗酸化活性の比較検討

    日本家政学会, 2007年

  • COX-2発現抑制及びPPAR活性化を指標にした植物油の機能性評価

    日本栄養食糧学会, 2007年

  • レスベラトロール関連物質による核内受容体PPARの活性化の検討

    日本栄養食糧学会, 2007年

  • 大腸菌を用いたミラクリンの発現と精製

    日本栄養食糧学会, 2007年

  • 葉酸欠乏ラットにおける遺伝子発現の変化

    日本栄養食糧学会, 2007年

  • Analysis of gene expression related to enrich environment after ischemic stroke in rats

    Brain’07 (The 23rd International symposium on Cerebral Blood Flow, Metabolism and Function), 2007年

  • 葉酸欠乏ラットによる遺伝子発現の変化

    日本ビタミン学会, 2007年

  • 植物ポリフェノール長期摂取による PPARαに依存した寿命延長効果の可能性

    日本ビタミン学会, 2007年

  • Resveratrol: a selective suppressor of COX-2 expression and an activator for PPARs

    10th International Conference on Bioactive Lipids in Cancer, Inflammation, and Related Diseases, 2007年

  • レスベラトロールの分子作用機構:COX-2発現抑制及びPPAR活性化を中心にして

    日本栄養食糧学会中国四国、近畿支部合同大会, 2007年

  • Activation of PPAR by resveratrol and other polyphenols Hiroyasu Inoue, Rieko Nakata, Shobu Namura

    ICPH2007, 2007年

  • 大腸菌による味覚修飾タンパク質ミラクリンの発現と精製

    日本生化学会, 2007年

  • COX-2及び核内受容体PPARへの影響を指標にした植物油の機能性評価

    日本生化学会, 2007年

  • レスベラトロール関連物質による核内受容体PPAR活性化と抗酸化活性の比較検討

    日本生化学会, 2007年

  • Analysis of gene expression related to enrich environment after ischemic stroke in rats

    Brain’07 (The 23rd International symposium on Cerebral Blood Flow, Metabolism and Function), 2007年

  • Resveratrol: a selective suppressor of COX-2 expression and an activator for PPARs

    10th International Conference on Bioactive Lipids in Cancer, Inflammation, and Related Diseases, 2007年

  • Activation of PPAR by resveratrol and other polyphenols Hiroyasu Inoue, Rieko Nakata, Shobu Namura

    ICPH2007, 2007年

  • Possible linkage between cyclooxygenase-2 and nuclear receptor PPAR

    第14回武田科学振興財団生命科学シンポジウム, 2006年

  • 生活習慣病予防を目指した遺伝子プロモーターの解析

    第3回 サプリメント研究会 (apiRAHS)(招待講演), 2006年

  • Relationship between cyclooxygenase-2 and nuclear receptor PPAR as a target against lifestyle-related diseases

    Symposium on Cell Signaling and Gene Regulation, 2006年

  • 植物ポリフェノールによるPPAR活性化の検討:フレンチパラドックスとの関連

    情報計算化学生物学会(CBI学会)第270回研究講演会, 2006年

  • 植物性ポリフェノールと核内受容体 PPAR の相互作用の検討

    日本ビタミン学会, 2006年

  • 植物油による誘導型シクロオキシゲナーゼ発現抑制の検討

    日本家政学会, 2006年

  • 赤ワインに含まれるポリフェノール・レスベラトロールの核内受容体PPARを介する生活習慣病予防効果

    秋田応用生命科学研究会, 2006年

  • フレンチパラドックスと核内受容体PPARの新しい接点

    日本栄養食糧学会, 2006年

  • 植物性ポリフェノールによる核内受容体PPARの活性化

    日本脂質生化学会, 2006年

  • 4回膜貫通型膜蛋白質5-リポキシゲナーゼ活性化タンパク質の精製と結晶化

    蛋白質科学会, 2006年

  • Expression and Purification of Proteins Related to Arachidonate Cascade for Development of Novel Drugs

    第4回ナノテクノロジー総合シンポジウム, 2006年

  • Possible linkage between cyclooxygenase-2 and nuclear receptor PPAR

    15th Takeda Life Science symposium, 2006年

  • Relationship between cyclooxygenase-2 and nuclear receptor PPAR as a target against lifestyle-related diseases

    Symposium on Cell Signaling and Gene Regulation, 2006年

  • 核内受容体を用いた食品の機能性評価

    第6回機能性食品開発研究会, 2005年

  • Possible connection between “French Paradox” and PPAR

    International Conference on the Bioscience of Lipids, 2005年

  • 誘導型シクロオキシゲナーゼと核内受容体PPARとの関係

    Takeda Genome Urology, 2005年

  • ラット局所脳虚血モデルにおける炎症関連遺伝子群の経時的発現変動

    日本ビタミン学会, 2005年

  • Possible connection between “French Paradox” and PPAR

    International Conference on the Bioscience of Lipids, 2005年

  • 核内受容体PPARαの活性化とその分子作用機構の検討

    日本ビタミン学会第56会大会 (2004), 2004年

  • 核内受容体PPAR は「フレンチパラドックス」に関与するか?

    第27回日本分子生物学会 (2004) (ワークショップ), 2004年

  • 赤ワインに含まれるポリフェノールの新しい分子作用機構

    日本家政学会近畿支部会 (2004), 2004年

  • 赤ワインに含まれるポリフェノール・Resveratrolによる核内受容体PPARαの活性化とその脳保護効果

    日本ビタミン学会第55回大会 (2003), 2003年

  • 核内受容体PPARを介する誘導型シクロオキシゲナーゼの発現調節に関する研究

    第54回日本ビタミン学会(学会奨励賞受賞講演), 2002年

  • CYCLOOXYGENASE-2 EXPRESSION BY FLUID SHEAR STRESS IN VASCULAR ENDOTHELIAL CELLS

    12th International Conference on Prostaglandins and Related Compounds, 2002年

  • DISTINCT CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BETWEEN GLUCOCORTICOID RECEPTOR AND PPARγ

    Fifth International Workshop on COX-2, 2002年

  • CYCLOOXYGENASE-2 EXPRESSION BY FLUID SHEAR STRESS IN VASCULAR ENDOTHELIAL CELLS

    12th International Conference on Prostaglandins and Related Compounds, 2002年

  • DISTINCT CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BETWEEN GLUCOCORTICOID RECEPTOR AND PPARγ

    Fifth International Workshop on COX-2, 2002年

  • FEEDBACK CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BY PPARγ

    Seventh International Congress on Platelet-Activating Factor and Lipid Mediators, 2001年

  • FEEDBACK CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BY PPARγ

    4th International Symposium on Molecular Medicine, 2001年

  • FEEDBACK CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BY PPARγ

    Seventh International Congress on Platelet-Activating Factor and Lipid Mediators, 2001年

  • FEEDBACK CONTROL OF CYCLOOXYGENASE-2 EXPRESSION BY PPARγ

    4th International Symposium on Molecular Medicine, 2001年

  • Transcriptional regulation of COX-2 gene mediated through PPARg

    11th International Conference on Prostaglandins and Related Compounds, 2000年

  • Transcriptional regulation of COX-2 gene mediated through PPARg

    11th International Conference on Prostaglandins and Related Compounds, 2000年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    Keystone Symposia on Molecular & Cellular Biology, Lipid Mediators, 1999年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    IBC's Industry Symposium on COX-2 Inhibitors (Invited lecture), 1999年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    Keystone Symposia on Molecular & Cellular Biology, Lipid Mediators, 1999年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    IBC's Industry Symposium on COX-2 Inhibitors (Invited lecture), 1999年

  • Regulation of Prostacyclin and Thromboxane Synthesis in Cardiovascular System

    COE International Symposium, Sensing and Signaling in the Cardiovascular System, 1998年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Macrophages and Endothelial Cells

    IBC's Industry Symposium on COX-2 Inhibitors, 1998年

  • Regulation of Prostacyclin and Thromboxane Synthesis in Cardiovascular System

    COE International Symposium, Sensing and Signaling in the Cardiovascular System, 1998年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Macrophages and Endothelial Cells

    IBC's Industry Symposium on COX-2 Inhibitors, 1998年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    Gordon Research Conference, Biochemistry of Lipid & Lipid Mediators, 1997年

  • Distinct Regulation of Cyclooxygenase-2 Gene Expression between Endothelial Cells and Macrophages

    Gordon Research Conference, Biochemistry of Lipid & Lipid Mediators, 1997年

  • Transcriptional Regulation of Human Prostaglandin Endoperoxide Synthase-2 Gene in U937 Cells: Promoter Region Responsible for the Induction by Lipopolysaccharide and the Suppression by Dexamethasone

    10th International Conference on Prostaglandins and Related Compounds, 1996年

  • Transcriptional Regulation of Human Prostaglandin Endoperoxide Synthase-2 Gene in U937 Cells: Promoter Region Responsible for the Induction by Lipopolysaccharide and the Suppression by Dexamethasone

    10th International Conference on Prostaglandins and Related Compounds, 1996年

  • Transcriptional regulation of huan prostaglandin-endoperoxide synthase-2 gene in vascular endothelial cells.

    4th International Conference on Eicosanoids & other bioactive lipids in cancer,inflammation & radiation injury, 1995年

  • Transcriptional regulation of huan prostaglandin-endoperoxide synthase-2 gene in vascular endothelial cells.

    4th International Conference on Eicosanoids & other bioactive lipids in cancer,inflammation & radiation injury, 1995年

  • Structure and transcriptional regulation of the human prostaglandin endoperoxide syntase 2 gene

    9th International Conference on Prostaglandins and Related Compounds, 1994年

  • Structure and transcriptional regulation of the human prostaglandin endoperoxide syntase 2 gene

    9th International Conference on Prostaglandins and Related Compounds, 1994年

所属学協会

  • 日本生化学会

  • 日本栄養食糧学会

  • 日本味と匂い学会

  • 日本ポリフェノール学会

  • 日本肥満学会

  • 日本分子生物学会

  • 日本ビタミン学会

  • 日本家政学会

  • 日本脂質生化学会

  • 脂溶性ビタミン総合研究委員会

Works(作品等)

  • シロイヌナズナを用いたミラクリンの分子機能の解明

    2014年03月 - 2016年03月

  • シロイヌナズナを用いたミラクリンの大量発現

    2010年04月 - 2013年03月

  • 食品素材の機能性評価

    2007年04月

  • 植物性ポリフェノール(レスベラトロールなど)の新しい作用機構の解明と利用

    2005年05月 - 2006年03月



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