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Effects of alteration in olasma membrane components on vascular endothelial cell function.

Research Project

Project/Area Number 07457035
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Pathological medical chemistry
Research InstitutionTokyo Medical and Dental University

Principal Investigator

MUROTA Sei-itsu  Tokyo Medical and Dental University, 歯学研究科, 教授 (50072989)

Co-Investigator(Kenkyū-buntansha) MORITA Ikuo  Tokyo Medical and Dental University, 歯学研究科, 助教授 (60100129)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1997: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1995: ¥4,400,000 (Direct Cost: ¥4,400,000)
KeywordsEndothelial cell / LDL-cholesterol / HMGCoA reductase / Simvastatin / membrane fluidity / EPA / HLGCoA還元酵素 / HMGCoA還元酵素
Research Abstract

theta-Toxin is a cholesterol-binding, pore-forming cytolysin of Clostridium perfringens. To detect cell surface cholerterol, we prepared a theta-toxin derivative, BCtheta by biotinylation of a protease-nicked theta-toxin, which has the same binding affinity for cholesterol as theta-toxin without cytolytic activity. Human erthyrocytes, V79 cells and human umbilical vein endothelial cells (HUVEC), were stained with BCtheta coupled with FITC-avidin, and then the cell were analyzed by either flow cytometry or laser confocal microscopy. Treatment of the cells with digitonin, a cholesterol-sequestering reagent, decreased the fluorescence intensity to the background level, indicating that BCtheta staining is specific for choresterol, The fluorescence intensity of erthrocytes pre-permeabilized with a small amount of theta-toxin increased more than ten-fold, suggesting higher cholesterol contents in the inner layer of the plasma membrane. When cells were cultured with cholestrol-depleted medium … More , the fluorescence intensity stained by BCtheta decreased remarkably in V79 cells, but did not change in HUVEC.This indicates that cell surface cholesterol may be provided in different ways with these two cell lines. These results suggest that BCtheta can be a useful probe for visualizing cell surface cholesterol and for evaluating the effects of cellular events on me topology and distribution of cholesterol. Next the membrane fluidity of the cholestrol-poor bovine carotid artery endothelial cells (BAEC) was examined. Chorestrol-poor BAEC were obtained by treating the cells with 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors under 10% low density lipoprotein (LDL)-deficient serum condition for 2 days. Simvastatin reduced the intracellular cholesterol content significantly at a concentration of 0.1 mug/ml. The reduction in the cholesterol content was accompanied by the enhancement of the cell membrane fluidity which was measured by a photobleaching technique. Additional data suggested that the reduction in cholesterol content referred to the reduction in the proliferation of BAEC.Both VEGF and bFGF activated mitogen-activated protein (MAP) kinase in BAE cells, however, EPA selectively inhibited VEGF-induced, but not bFGF-induced activation of MAP kinase. Flk-1 expression was inhibited dose-dependently in EPA-treated cells whilst Flt-1 expression was increased in EPA treated cells. This in vitro inhibitory effect by EPA on Flk-l receptor expression provides indirect evidence that one of the mechanisms of EPA for anti-tumor action in vivo may be related to its anti-angiogenic action. Less

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] I.MORITA I.SATO,L.MA and S.MUROTA: "Enhancement of membrane fluidity in choresterole-poor endo-thelial cells pre-treated with simvastatin." ENDOTHELIUM. 5. 107-113 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.IWAMOTO,I.MORITA,M.FUKUDA,S.MUROTA,S.ANDO and Y.OHNO IWASHITA: "A biotinylated perfrigolysin O derivative:A new probe for detection of cell surface choresterol." BIOCHIM.BIOPHYS.ACTA.,. 1327. 222-230 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] I.SATO,L.MA,IKEDA,I.MORITA and S.MUROTA: "Simvastatin, a potent HMG-CoA reductase inhibitor, inhibits the proliferation of human and bovine endothelial cells in vitro." J.ATHEROSCR.THROMB.,. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.YANG,I.MORITA and S.MUROTA: "Eicosapentaenoic acid attenuates vascular endothelial growth factor-induced proliferation via inhibiting FLK-1 receptor expression in bovine caroted artery endotehlial cells" J.CELL.PHYSIOL.,. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 〓IRIE,F.TSUKAHARA,E.FUJIE,Y.UCHIDA,YOSHIOKA,W.R.HE,M.SHITASHIGE,S.MUROTA,T.MURAKI: "Cationic amino acid transporter-2 mRNA induction by tumor necrosis factor-a in vascular endotehlial cells." EUR.J.PHARMAC.,. 339. 289-293 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] KAMEDA,I.MORITA,M.HANDA,J.KABURAKI,YOSHIDA,T.MIMORI,S.MUROTA,and I.IKEDA: "Re-expression of functional P-selectin molecules on the 〓〓do-thelial cell surfase choresterol." BRI.J.HAEMATOL.,. 97. 348-355 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 〓ORITA,M.SHINDLER,D.DEWLITT,S.MUROTA,and W.SMITH: "Eicosanoids and other bioactive lipids in cancer, infla-mmation, Andradiationinjury 3" A novel method for prostaglandin endoperoxide H synthase activity in individual intact cells., 521-524 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.MUROTA,M.ONODA,S.YANG,Y.CHANG,I.SATO SUZUKI,T.KANAYASU TOYODA,and S.MUROTA: "Eicosanoids and vascular endothelial cell function." Proceedings of 4th international congress on Fatty Acid and Eicosanoids (in press),

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] I.Morita, I.Sato, L.Ma and S,Murota: "Enhancement of membrane fluidity in choresterole-poor endothelial cells pre-treated with simvastatin." Endothelium. 5. 107-113 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Iwamoto, I.Morita, M.Fukuda, S.Murota and S.Ando: "A buotinylated perfrigolysin O derivative : A new probe for detection ofcell surface choresterol." Biochim.Biophys.Acta.1327. 222-230 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] I.Sato, L.Ma, M.Ikeda, I.Morita and S.Murota: "Simvastatin, a potent HMG-CoA reductase inhibitor, inhubits the proliferation of human and bovine endothelial cells in vitro." J.Atheroroscr.Thromb.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S,Yang, I.Morita and S.Murota: "Eicosapentaenoic acid attenuates vascular endothelial growth factor-induced proliferation via inhibiting FLK-1 receptor expression in bovine caroted artery endothelial cells." J.Cell.Physiol.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Irie, F.Tsukahara, E.Fujie S.Murota et al.: "Cationic amino acid transporter-2 MRNA induction by tumor necrosis factor-a in vascular endothelial cells." Eur.J.Pharmac.339. 289-293 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Kameda, I.Morita, S.Murota et al.: "Re-expression of functional P-selectin molecules on the endothelial cell surfase choreterol." Bri.J.Haematol.97. 348-355 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] I.Sato-Suzuki and S.Murota: "Simvastatin inhibits the division and induces neurite-like outgrouth in PC12 cells." Neuroscle.Letters. 220. 21-24 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] S.Sano, I.Sato-Suzuki, H.Fujita, I.Morita, M.Nagao, S.Murota: "NO is not involved in the simvastatin induced cell division and differentiation in PC12 cells." Neurosci.Letters. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] I.MORITA, I.SATO, L.MA and S.MUROTA: "Enhancement of membrane fluidity in choresterole-poor endothelial cells pre-treated with simvastatin." ENDOTEHLIUM. 5. 107-113 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.KAMEDA, I.MORITA, M.HANDA, J.KABURAKI, T.YOSHIDA, T.MIMORI,S.MUROTA and Y.IKEDA: "Re-expression of functional P-selectin molecules on the endothelis cell surface by repeated stimulation with thrombin." BRL.J.HAEMATOL.97. 348-355 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] IWAMOTO, I.MORITA, M.FUKUDA, MUROTA, S.ANDO and Y.OHNO-IWASHITA: "A biotinylated perfrigolysin O derivative:A new probe for detectic of cell surface choresterol." BIOCHIM.BIOPHYS.ACTA.1327. 222-230 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.SHITASHIGE, I.MORITA and S.MUROTA: "Different sustance utilization between prostaglandin endoperoxide H synthase-1 and -2 in NIH3T3 fibroblasts." BIOCHIM.BIOPHYS.ACTA.1389. 57-66 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] I.SATO, L.MA, M.IKEDA, I.MORITA and S.MUROTA: "Simvastatin,a potent HMG-CoA reductase inhibitor,inhibits the proliferation of human and bovine endothelial cells in vitro." J.ATHROSCR.THROMB.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] S.YANG, I.MORITA and S.MUROTA: "Eicosapentaenoic acid attenuates vascular endothelial growth factor-induced proliferation via inhibiting FLK-1 receptor expression in bovine carotid artery endothelial cells." J.CELL.PHYSIOL.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] I.MORITA, M.SHINDLER, D.DEWITT, S.MUROTA, W.SMITH: "Eicosanoids and other bioactive lipids in cancer,inflammation andradiationinjury3" A novel method for prostaglandin endoperoxide H synthase activity in individual intact cells., 521-524 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] MUROTA, M.ONODERA, S.YANG, Y.CHANG, SATO-SUZUKI, T.KANAYASU-TOYODA: "Eicosanoids and vascular endothelial cell functions." Proceedings of 4th international congress on Essential Fatty Acids and Eicosanoids(in press),

    • Related Report
      1997 Annual Research Report
  • [Publications] I.Morita,I.Sato,L.Ma and S.Murota: "Enhancement of Membrane Fluidity in Choresterol-Poor Endothelial Cells Pre-treated with Simvastatin" Endothelium. (in press).

    • Related Report
      1996 Annual Research Report
  • [Publications] I.Sato and S.Murota: "Paracrine function of endothelium-delived nitric oxide." LIFE SCI.,. 56. 1079-1087 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Y.Wakabayashi,H.Fujita,I.Morita,K.Kawaguchi and S.Murota: "Conversion of xanthine oxidase in bovine caroid artery endothelial cells induced activated neutrophils:Involvement of adhesion molecules." BIOCHIM.BIOPHYS.ACTA.,. 1265. 103-109 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] I.Sato,K.Kaji and S.Murota: "Age related decline in cytokine induced nitric oxide synthase activation and apoptosis in cultured endothelial cells:minimal involvement of nitric oxide in the apoptosis." MECH.AGING.DEV.,. 81. 27-36 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] S.Murota,H.Fujita,I.Morita and Y.Wakabayashi: "ANNALS OF THE NEW YORK ACADEMY OF SCIENCES,RECENT ADVANCES IN ATHEROSCLEROSIS RESEARCH" The New York Academy of Sciences., 133-147 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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