• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Free Radical Generation and Cell Injury in Endothelial Cell Co-Incubated with Neutrophil

Research Project

Project/Area Number 01570483
Research Category

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

Allocation TypeSingle-year Grants
Research Field Circulatory organs internal medicine
Research InstitutionOsaka University

Principal Investigator

KUZUYA Tsunehiko  Osaka University, School of Medicine, Assistant Professor, 医学部, 助手 (80150340)

Co-Investigator(Kenkyū-buntansha) INUI Makoto  Osaka University, School of Medicine, Assistant Professor, 医学部, 助手 (70223237)
TADA Michihiko  Osaka University, School of Medicine, Professor, 医学部, 教授 (90093434)
星田 四朗  大阪大学, 医学部附属病院, 医員
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1990: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1989: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsNeutrophil / Endothelial Cell / Myocardial Cell / Cell Injury / Adhesion / Oxygen-Free Radicals / リボキシゲナ-ゼ代謝 / 粘着
Research Abstract

Activated neurtophils play an important role in myocardial injury during ischemia followed by reperfusion. To clarify the mechanism of the activation of neutrophils in the reperfused microvasculature, we co-incubated canine heart capillary endothelial cells and isolated neutrophils under hypoxia-reoxygenation. The extent of neutrophil adhesion was evaluated by counting of cells which were adhered to the endothelial cell monolayer. Production of an arachidonate lipoxygenase metabolite, hydroxyeicosa-tetraenoic acid (HETE), was assayed by HPLC. Free radical generation was assayed by luminol-enhanced chemiluminescence and 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO) spin trapping of electron spin resonance (ESR) spectroscopy. After reoxygenation, serial changes of free radical generation revealed two peaks of increase ; the first was at 1 hour and the second was at 3 hours after reoxygenation. The first peak was diminished by allopurinol, whereas the second was diminished by nordihydroguaiare … More tic acid. Production of 15-, 5-, and 12-HETEs were augmented in endothelial cells after reoxygenation. Adhesion of neutrophils co-incubated with reoxygenated endothelial cells was also enhanced compared with that of neutrophils co-incubated with control endothelial cells. Adhesion of neutrophils was associated with the production of HETEs and free radicals at 3 hours after reoxygenation.
To elucidate whether these activated neutrophils produce myocardial cell injury, we co-cultured endothelial cells and myocardial cells in double chamber (myocytes on dish bottom ; lower chamber, endothelial cells on collagen sheet ; upper chamber) under hypoxia-reoxygenation. After reoxygenation, neutrophils were added to these cells and serial changes of free radical production was assayed by ESR and cell injury was determined by morphological change. Under mono-culture of endothelial cells, DMPO-OH signal was detected mainly at the late phase of reoxygenation, while DMPO-OOH signal was detected at the early phase of reoxygenation. Under the co-culture condition, DMPO-OOH signal was markedly enhanced in the upper chamber, compared with the sum of that generated in mono-culture. DMPO-OH signal was also enhanced after the late phase of reoxygenation. Injury of myocardial cells and endothelial cells was augmented in a close relation with the enhancement of free radical generation. These results suggested that reoxygenation injury of endothelial cells induced neutrophil activation and augmentation of free radical production, resulting in the enhancement of myocardial cell injury. Less

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Kuzuya,T.et al.: "Role of free radicals and neutrophils in.canine myocardial reperfusion injury:myocardial salvage by a novel free radical scavenger,2ーoctaーdecylascorbic acid." Cardiovasc.Res. 23. 323-330 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida,S.et al.: "Attenuation of neutrophil function by inhibitors of arachidonate metabolism reduces the extent of canine myocardial infarction." Am.J.Cardiol. 63. 24E-28E (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nishida,.et al: "Polymorphcnuclear leukocytes induced vasoconstriction in isolated canine coronary arteries." Circ Res.66. 253-258 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kuzuya T.et al.: "Detection of oxygenーderived free radical generation in the canine postischemic heart during late phase of reperfusion." Circ Res.66. 1160-1165 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida,S.et al: "Relationship between leukocyte activity and arrhythmias in acute myocardial infarction." Cardiology Board Review. 7. 48-60 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida,S.et al: "Transcardiac alteration of neutrophil function before and after coronary thrombolysis in human myocardial infarction." Int.J.Cardiol.30. 49-54 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nishida,M.et al: "Elservier Science Pubishers B.V.,Amusterdam." In Medical Biochemical and Chemical Aspects of Free Radicals, 1097-1100 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida,S.et al: "Elservier Science Publishers B.V.,Amsterdam." In Medical Biochemical and Chemical Aspects of Free Radicals, 489-492 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kuzuya T. Hoshida S. Nishida M. Kim Y. Fuji H. Kitabatake A. Kamada T. Tada M.: "Role of free radicals and neutrophils in canine myocardial reperfusion injury : myocardial salvage by a novel free radical scavenger, 2-octa-decylascorbic acid," Cardiovasc. Res. 23. 323-330 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida S. Kuzuya T. Nishida M. Kim Y. Kitabatake A. Kamada T. Tada M.: "Attenuation of neutrophil function by inhibitors of arachidonate metabolism reduces the extent of canine myocardial infarction." Am. J. Cardiol.63. 24E-28E (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nishida M. Kuzuya T. Hoshida S. Kim Y. Fuji H. Kitabatake A. Kamada T. Tada M.: "Polymorphonuclear leukocytes induced vasoconstriction in isolated canine coronary arteries." Circ Res.66. 253-258 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kuzuya T. Hoshida S. Kim Y. Nishida M. Fuji H. Kitabatake A. Tada M. Kamada T.: "Detection of oxygen-derived free radical generation in the canine post-ischemic heart during late phase of reperfusion." Circ Res.66. 1160-1165 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida S. Kuzuya T. Kamada T. Tada M.: "Relationship between leukocyte activity and arrhythmias in acute myocardial infarction." Cardiology Board Review. 7. 48-60 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida S. Kuzuya T. Nishida M. Kim Y. Fuji H. Minamino T. Kitabatake A. Tada M. Kamada T.: "Transcardiac alteration of neutrophil function before and after coronary thrombolysis in human myocardial infarction." Int. J. Cardiol. 30. 49-54 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nishida M. Kazuya T. Hoshida S. Kim Y. Fuji H. Kitabatake A. Kamada T. Tada M.: "The effect of endothelial cells on superoxide anion and arachidonate lipoxygenase metabolism of leukocytes. In Medical. Biochemical and Chemical Aspects of Free Radicals (Hayaishi O. Niki E. Kondo M. Yoshikawa T.)" Elservier Science Publishers B. V., Amsterdam. 1097-1100 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hoshida S. Kuzuya T. Nishida M. Kim Y. Fuji H. Li Y-W, Kitabatake A. Kamada T. Tada M.: "Reactive oxygen species and arachidonate metabolism in luminol-enhance chemiluminescence of activated neutrophils. In Medical. Biochemical and Chemical Aspects of Free Radicals (Hayaishi O. Niki E. Kondo M. Yoshikawa T.)" Elservier Science Publishers B. V., Amsterdam. 489-492 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nishida,M.;et al: "Polymorphonuclear leukocytes induced vasoconstriction in isolated canine coronary arteries." Circ Res.66. 253-258 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kuzuya,T.;et al: "Detection of oxygenーderived free radical generation in the canine postischemic heart late phase of reperfusion." Circ Res.66. 1160-1165 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hoshida,S;et al: "Relationship between leukocyte activity and arrhythmias in acute myocardial infarction." Cardiology Board Review. 7. 48-60 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] T.Kuzuya et al.: "Role of free radicals and neutrophils incanine myocardial reperfusion injury: myocardial salvage by a novel free radical scavenger,2ーoctadecylascorbic acid." Cardiovasc.Res.23. 323-330 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] S.Hoshida et al.: "Attenuation of neutrophil function by inhibtors of arachidonate metabolism reduces the extent of canine myocardial infarction." Am.J.Cardiol.63. 24E-28E (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] M.Nishida et al.: "Polymorphonuclear leukocytes induced vasoconstriction in isolated canine coronary arteries." Circ.Res.66. 253-258 (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] T.Kuzuya et al.: "Detection of oxygenーderived free radical generation in the canine postischemic heart during late phase of reperfusion." Circ.Res.66. (1990)

    • Related Report
      1989 Annual Research Report

URL: 

Published: 1989-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi