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Cytotoxicity of 4-hydroxynonenal enantiomers and stereoselectivity in those detoxification metabolism

Research Project

Project/Area Number 14572071
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionTokyo University of Pharmacy and Life Sciences

Principal Investigator

HIRATSUKA Akira  Tokyo University of Pharmacy and Life Sciences, School of Pharmacy, Professor, 薬学部, 教授 (20165179)

Project Period (FY) 2002 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Keywords4-hydroxynonenal / Clone 9 / cell toxicity / glutathione S-transferase / hydroperoxide / stereoselectivity / metabolism / detoxification / エナンチオマー / 立体選択性 / グルタチオンS-トランスフェラーゼ / グルタチオン抱合 / 還元反応 / 細胞毒性 / PC12 / ラット / 脂質過酸化 / Clone 9
Research Abstract

Racemic 4-hydroxy-2(E)-nonenal (HNE) is known as a reactive and highly toxic product released from biomembranes by lipid peroxidation in vivo. Until the present, all previous studies on the cytotoxic effects of HNE were performed using its racemate. In this study, the toxicities of (S)- and (R)-HNE were examined in rat Clone 9 cells. IC_<50> values for inhibition of cell growth were 15.6 μM for (R)-HNE and 5.1 μM for (S)-HNE. To estimate the cytotoxic response, Clone 9 cells were separately incubated with (S)- and (R)-HNE, stained with Annexine V-FITC or propidium iodide, and analyzed by flow cytometry. These results suggest that (S)-HNE is a much more potent agent than (R)-HNE for cell growth inhibition and induction of apoptosis and also that cell death via apoptosis and via necrosis occurs only at lower (<10 μM) and higher (>20 μM) concentrations of HNEs, respectively. This study also showed that in rat Clone 9 cell cytosol, FINE enantiomers were detoxified (S)-preferentially by GSH conjugation and that several classes of GST existed in rat Clone 9 cells. However, no immunostainable protein was detected in the cell cytosol by Western blot analysis using the anti-rGSTA4-4 antibody. The absence of the rGSTA4-4 isoform could be responsible for the observed low Vmax/Km value in the GSH conjugation of HNE enantiomers in Clone 9 cell cytosol compared with a high Vmax/Km value observed for that in rat liver cytosol. The cytotoxic study in rat Clone 9 cells revealed that (S)-HNE displayed significant toxicity in the cell line studied, with the IC_<50> value in the low micromolar range. Thus, based on the results presented here, enantiopreferential detoxification of HNE enantiomers in Clone 9 cells is unlikely to be an important factor in determining their response to (S)-preferential cytotoxicity.

Report

(5 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (4 results)

All 2004 2003 Other

All Journal Article (3 results) Publications (1 results)

  • [Journal Article] The induction mechanism of glutathione transferase by ultraviolet light, and its toxicological significance2004

    • Author(s)
      Akira Hiratsuka
    • Journal Title

      Environmental Mutagen Research 26

      Pages: 107-115

    • NAID

      10013956800

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 紫外線によるグルタチオン転移酵素の誘導機構とその毒性学的意義2004

    • Author(s)
      平塚 明
    • Journal Title

      Environ.Mutagen.Res. 26

      Pages: 107-107

    • NAID

      10013956800

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Clinical implications of the genetic polymorphisms in Phase II drug-metabolizing enzymes and dihydropyrimidine dehydrogenase2003

    • Author(s)
      Akira Hiratsuka
    • Journal Title

      Japanese Journal of Clinical Pharmacology and Therapeutics 34(4)

      Pages: 163-170

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Publications] 平塚 明: "薬物代謝第2相酵素およびジヒドロピリミジンデヒドロゲナーゼの遺伝多型の臨床的関わり"Jpn.J.Clin.Pharmacol.Ther.. 34・4. 163-170 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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