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Study on mechanism of phospholipae A_2 activation.

Research Project

Project/Area Number 63580159
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionKansai Medical University

Principal Investigator

SUGATANI Junko  Kansai Medical, University, Department of Medical, Chemistry, Instructor, 医化学教室, 講師 (30098131)

Project Period (FY) 1988 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordsphospholipase A_2 / phospholipase A_2 inhibitory protein / ionophore A23187 / platelet activation / arachidonic acid release / platelet-activating factor / gastric PAF / scintillation proximity radioimmunoassay / GC@MS / シンチレ-ション・プロキシミティ-・ラジオイムノアッセイ(SPRIA) / ホスホリパーゼA_2 / ホスホリパーゼA_2阻害因子 / 血小板活性化因子(platelet-activating factor,PAF) / PAFアセチルハイドロラーゼ
Research Abstract

(1) Platelet phospholipae A_2 inhibitory protein. 1-Acyl-2-[^<14>4C]arachidonyl GPC was not hydrolyzed by beef platelet lysate (freeze-thawing platelet)(as an enzyme) in the presence of 10 mM Ca^<2+>. The addition of 10 mM Ca^<2+> to [^3H]arachidonic acid-labelled platelet membrane made [^3H]-arachidonic acid released, but the membrane-bound phospholipase A_2 activity was suppressed by beef platelet lysate even in the presence of 10 mM Ca^<2+>. We elucidated the presence of platelet phospholipase A_2 inhibitory protein in beef platelet, which was distinct from lipocortin.
(2) Distinct mechanism of ionophore A23187-induced platelet activation in the presence and absence of extracellular Ca^<2+>. In order to study the mechanism of platelet phospholipase A_2 activation, we investigated A23187-induced platelet activation using washed rabbit platelets double-labelled with [^<32>P]phosphoric acid and [^3H]arachidonic acid in the presence and absence of extracellular Ca^<2+>. In the absence of … More extracellular Ca^<2+>, A23187 increased the amount of intracellular Ca^<2+> which was lower than that in the presence of extracellular Ca^<2+>. TPA induced 40 kDa and 20 kDa protein phosphorylation, regardless of extracellular Ca^<2+>. In TPA-treated platelets, A23187-induced arachidonic acid release increased in both the presence and absence of extracellular Ca^<2+>, but inositol phospholipid metabolism was not affected by A23187 in the absence of extracellular Ca^<2+>. We elucidated thus distinct mechanism of A23187-induced platelet activation in the presence and absence of extracellular Ca^<2+>.
(3) Occurrence of platelet-activating factor (PAF) in normal rat stomach and alteration of PA level by water-immersion stress. In order to elucidate the mechanism of PAF biosynthesis connected with phospholipase A_2 activation, we detected platelet-activating substance in gastrointestinal areas, which was confirmed by GC/MS analysis. In the normal rat stomach, the level of PAF was high in the antral mucosa. The percentage composition of each molecular species of PAF was 16:OPAF (34%) and 18:OPAF (66%). Application of water-immersion stress affected not only the amounts of PAF but also their molecular heterogeneity in the glandular stomach. The amount of 18:OPAF increased markedly (to 4-fold) in the corpus along with severe lesions after stress for 7 h. These changes might be associated with the pathogenicity of gastric ulcer.
(4) Development of a novel scintillation proximity radioimmunoassay for platelet-activating factor measurement. The SPRIA assay system was suitable for the quantitation of 0.03 to 2 pmol of 16:OPAF. The cross-reactivity was high with 1- alkyl-2-acetyl GPC but was very low with PAF analogs and PAF antagonists. The specificity of SPRIA was higher than that of bioassay, quite different from that of the platelet receptor. Less

Report

(3 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] 菅谷純子: "Platelet-activating factorと血管病変" 臨床科学. 24. 1220-1228 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 三輪匡男: "気管支喘息児童血清中のPAFアセチルハイドロラ-ゼ活性" 炎症. 8. 327-333 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Masao Miwa: "Characterization of serum platelet-activating factor(PAF)acetylhydrolase:Correlation between deficiency of serum PAF acetylhydrolase and respiratory symptoms in asthmatic children." J.Clin.Invest.82. 1983-1991 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Occurrence of platelet-activating factor(PAF)in normal rat stomach and alteration of PAF level by water immerision stress." FASEB J.3. 65-70 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 菅谷純子: "ホスホリパ-ゼ" 総合脂質科学 鹿山光編 恒星社厚生閣. 385-400 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 菅谷純子: "PAFの細胞生物学:血小板" 現代化学 増刊17 血小板活性化因子ー生化学・生理・病理 井上圭三,和久敬三編 東京化学同人. 100-107 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Kazuyo Fujimura: "Serum platelet-activating factor acetylhydrolase activity in rats with gastric ulcers induced by water-immersion stress." Scand.J.Gastroenterol.24(Suppl 162). 59-62 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 菅谷純子: "PAFによる血小板活性化とその病態生理学的役割" 最新医学. 45. 486-491 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Development of a novel scintillation protimity radioimmunoassay for platelet-activating factor measurement:comparison with bioassay and GC/MS techniques" Life Sciences. (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Molecular heterogeneity of platelet-activating factor(PAF)in normal rat glandular stomach determined by gas chromatography/mass spectrometry:" Lipids. (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Masao Miwa: "Characterization of serum platelet-activating factor (PAF) acetyl-hydrolase : Correlation between deficiency of serum PAF acetyl-hydrolase and respiratory symptoms in asthmatic children." J. Clin. Invest. 82, 1983-1991(1988).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Occurrence of platelet-activating factor (PAF) in normal rat stomach and alteration of PAF level by water immersion stress." FASEB J. 3, 65-70(1989).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Kazuyo Fujimura: "Serum platelet-activating factor acetyl-hydrolase activity in rats with gastric ulcers induced by water-immersion stress." Scand. J. Gastroenterol. 24(suppl 162), 59-62(1989).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Development of a novel scintillation proximity radioimmunoassay for platelet-activating factor measurement: comparison with bioassay and GC/MS techniques" Life Sciences (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Junko Sugatani: "Molecular heterogeneity of platelet-activating factor (PAF) in normal rat glandular stomach determined by gas chromatography/mass spectrometry: alteration in molecular species of PAF by water-immersion stress." Lipids (1990).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 菅谷純子: "ホスホリパ-ゼ" 総合脂質科学 鹿山光編 恒星者厚生閣. 385-400 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 菅谷純子: "PAFの細胞生物学:血小板" 現代科学増刊17 血小板活性化因子-生化学・生理病理 井上圭三,和久敬三編 東京化学同人. 100-107 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kazuyo Fujimura: "Serum platelet-activating factor acetyl hydrolase activity in rats with gastric ulcers induced.by water-immersion stress." Scand.J.Gastro enterol.24(suppl 162). 59-62 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 菅谷純子: "PAFによる血小板活性化とその病態生理学的役割" 最新医学. 45. 486-491 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Junko Sugatani: "Development of a novel scintillation proximity radioimmunoassay for platelet-activating factor measurement:comparison with bioassay and GC/ML techniques" Life Sciences. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Junko Sugatani: "Molecular heterogeneity of platelet-activating factor(PAF)in nornal ret galndular stomach determined by gas chromatography/mass spectrometry:alteration in molecular species of PAF by water-immersion stress." Lipids. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 菅谷純子: 臨床科学. 24. 1220-1228 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Masao Miwa: J.Clin.Invest. 82. 1983-1991 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Junko Sugatani: FASEB J.3. 65-70 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Kazuyo Fujimura: Scand.J.Gastroenterol.Supplement. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] 菅谷純子: 現代化学増刊号:血小板活性化因子 (PAF)生化学・生理・病理. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] 菅谷純子: "総合脂質科学 分担:ホスホリパーゼ" 鹿山光 編、恒星社厚生閣, 385-400 (1989)

    • Related Report
      1988 Annual Research Report

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Published: 1988-04-01   Modified: 2016-04-21  

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