化学発光測定法によるBile Salt Export Pump輸送活性の測定  [in Japanese] Measurement of the Transport Activities of Bile Salt Export Pump Using Chemiluminescence Detection Method  [in Japanese]

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Author(s)

    • 山口 佳奈 YAMAGUCHI Kana
    • 北海道医療大学薬学部生命物理科学講座薬品分析化学教室 Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
    • 村井 毅 MURAI Tsuyoshi
    • 北海道医療大学薬学部生命物理科学講座薬品分析化学教室 Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
    • 惠 淑萍 HUI Shu-Ping
    • 北海道医療大学薬学部生命物理科学講座薬品分析化学教室 Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido
    • 黒澤 隆夫 KUROSAWA Takao
    • 北海道医療大学薬学部生命物理科学講座薬品分析化学教室 Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido

Abstract

  Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI)-labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3α-hydroxysteroid dehydrogenase (3α-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3α-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.<br>

Journal

  • YAKUGAKU ZASSHI

    YAKUGAKU ZASSHI 130(5), 755-761, 2010

    The Pharmaceutical Society of Japan

Codes

  • NII Article ID (NAID)
    130000259466
  • NII NACSIS-CAT ID (NCID)
    AN00284903
  • Text Lang
    JPN
  • ISSN
    0031-6903
  • NDL Article ID
    10697804
  • NDL Source Classification
    ZS51(科学技術--薬学)
  • NDL Call No.
    Z19-411
  • Data Source
    NDL  J-STAGE 
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