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Immunohistochemical study of cytokine induction and endotherial cell damage on hypoxic brain injury

Research Project

Project/Area Number 13671141
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Embryonic/Neonatal medicine
Research InstitutionKyoto Prefectural University of Medicine

Principal Investigator

HASEGAWA Koh  Kyoto Prefectural University of Medicine, Department of Medical Research, Associate Professor, 医学研究科, 講師 (80228444)

Co-Investigator(Kenkyū-buntansha) 吉岡 博  京都府立医科大学, 医学部, 助教授 (50128724)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥500,000 (Direct Cost: ¥500,000)
Keywordshypoxic brain injury / cytokine / nitric oxide / nitrotyrosine / free radical scavenger / 血管内皮細胞
Research Abstract

It is reported that Cytokines play important roles on hypoxic ischemic brain injury. In this study, the relationship between cytokines and nitric oxide (NO) in brain injury was investigated.
[Material and Method]
In each experiment forty cerebrocortical slices, each 350μm thick, were obtained from ten 7-day old Sprague-Dawley rats. Perfused slices had only one injury layer, were allowed 3 hrs to metabolically recover from decapitation, and were studied in a tissue culture well. In each study there was a 30 mm treatment period, such as hypoxia or NMDA, followed by 4 hours of perfusion with oxygenated ACSF. Frozen sections 20 μm thick were cut in the vertical plane of slices, and processed for Immunohistochemistry of IL1-beta. Adjacent frozen sections were used for HE staining and TUNEL staining. Expression of nitrotyrosine (NT), a marker of peroxynitrite was also investigated with/without 7-nitroindazole (7NI), a novel nNOS inhibitor. Apoptosis in slices was detected by TUNEL staining after etarabon (MCI-186) treatment before NMDA perfusion, a kind of free radical scavenger.
[Results]
Hypoxia or NMDA treatment caused IL1-beta expression at 15 to 30 min after the each insult, compared to control slices. 7NI could not decrease neuronal damage and NT positive cells, showing that NO did not cause neuronal death at early phase after hypoxia. Etarabon significantly decreased TUNEL positive cells on brain slices after NMDA treatment.
[Conclusion]
It is suggested that cytokine, which appears in neuronal tissue just after hypoxia, might cause NO induction by activation of the NMDA receptor, resulting in apoptosis of neuronal cells appeared at late phase.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 長谷川 功: "神経細胞死におけるNOの関与-脳スライス灌流モデルによる検討-"最新医学. 56. 498-502 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Minako Kihara: "Stimulation of NMDA receptors inhibits neuronal migration I embryonic cerebral cortex: a tissue culture study"Developmental Brain Research. 138. 195-198 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masafumi Morimoto: "Infantile spasms in a patient with William syndrome and craniosynostosis"Epilepsia. 44. 1459-1462 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Koh Hasegawa: "Neuronal death and NO in respiring cerebrocortical slices"Saishin-Igaku. 56(2). 498-502 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Minako Kihara: "Stimulation of NMDA receptors inhibits neuronal migration I embryonic cerebral cortex : a tissue culture study"Developmental Brain Research. 138. 195-198 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masafumi Morimoto: "Infantile spasms in a patient with William syndrome and craniosynostosis"Epirepsia. 44. 1459-1462 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Masafumi Morimoto: "Infantile spasms in a patient with William syndrome and craniosynostosis"Epilepsia. 44. 1459-1462 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Minako Kihara: "Stimulation of N-methyl-D-aspartate (NMDA) receptors inhibits neuronal migration in embryonic cerebral cortx : a tissue culture study"Developmental Brain Research. 138. 195-198 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 長谷川 功: "神経細胞死におけるNOの関与 脳スライス灌流モデルによる検討"最新医学. 53(3). 498-502 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 長谷川 功: "最新NICUマニュアル 第2版"診断と治療社. 233 (2002)

    • Related Report
      2001 Annual Research Report

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

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