Molecular mechanisms of neuronal death in ischemia

缺血时神经元死亡的分子机制

• 批准号: 05671158
• 负责人: NOZAKI Kazuhiko
• 金额: $ 1.47万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671158/
• 关键词: Ischemia; Apoptosis; delayd neuronal death; 神経細胞孔; 分子生物学; ストレス蛋白質; 成長因子; 神経細胞死

In order to elucidate the mechanisms of neuronal death in ischemic brain, we investigated the occurrence of DNA fragmentation in gerbil forebrain ischemia, and evaluated the effects of several specific drugs on DNA fragmentation. Forebrain ischemia was produced by transient occlusion of bilateral common carotid arteries for 10 minutes, and DNA fragmentation was confirmed using TUNEL method on cerebral cortex, caudate-putamen, and CA1 region of hippocampus. Administration of Cycloheximide (protein synthesis inhibitor) and aurintricarboxylic acid (endonuclease inhibitor) blocked the occurrence of DNA fragmentation completely in cerebral cortex and partially in caudate-putamen and hippocampus. Similar effects were observed by the administration of GYKI52466 (non-NMDA receptor inhibitor) and leupeptin (calpain inhibitor), but the administration of MK-801 (NMDA receptor inhibitor) showed no blocking effects. 7 nitro-indazole (NOS inhibitor) did show only minor blocking effects and FK 506 showed marked blocking effects.In order to examine the involvement of ICE family, a family of apoptosis-related gene, on ischemic neuronal death, the expression ICE and YAMA gene was measured by northern blotting after rat focal ischemia. The amount of ICE mRNA did not change after ischemia, but Messenger RNA of YAMA was markedly increased 10-20 hours after ischemia.We are now producing the reproduciblc mouse ischemic model to evaluate the involvement of apotosis-related gene on ishemic neuronal death.

为了阐明缺血性大脑中神经元死亡的机制，我们研究了沙鼠前脑缺血中DNA碎片的发生，并评估了几种特定药物对DNA碎片化的影响。前脑缺血是通过双侧共同的颈动脉暂时闭塞10分钟产生的，并使用TUNEL方法在脑皮层，尾状甲状腺素和海马的CA1区域确认DNA片段化。施用环己酰亚胺（蛋白质合成抑制剂）和甲酸羧酸（核酸内切酶抑制剂）完全阻断了大脑皮层中DNA碎片的发生，部分在尾状甲状腺素和海马中部分阻止了DNA碎片。通过给药GYKI52466（非NMDA受体抑制剂）和亮肽素（Calpain抑制剂）观察到了类似的影响，但是MK-801（NMDA受体抑制剂）的施用显示没有阻塞效应。 7硝基 - 诱导剂（NOS抑制剂）确实仅显示出较小的阻塞效应，FK 506显示出明显的阻塞效应。缺血后，冰mRNA的量没有变化，但是在缺血后10-20小时明显增加了YAMA的Messenger RNA。我们现在正在产生repoduciblc小鼠缺血模型，以评估与凋亡相关基因在等性神经元死亡上的参与。

项目成果

Shogo Nishi: "Ischemic tolerance due to the induction of HSP70 in" Brain Research. 615. 281-288 (1993)

Shogo Nishi：“脑研究中由于 HSP70 的诱导而产生缺血耐受性”。

T.Tokime et al: "Neuro protective effect of FK506 on transient global ischemia in gerbil" Neuro science Letters. (in press).

T.Tokime 等人：“FK506 对沙鼠短暂性整体缺血的神经保护作用”神经科学快报。

Koji Iihara: "Ischeinia induces the expresston of PDGF-B chain" Journal of Cerebral Blcod Flow Metab. 14. 818-824 (1994)

Koji Iihara：“Ischeinia 诱导 PDGF-B 链的表达”Journal of Cerebral Blcod Flow Metab。

T. Tokime: "Neuroprotective effect of FK506 on transient Shobal ischemia in gerbil." Neuroscience Letters. (in press).

T. Tokime：“FK506 对沙鼠短暂 Shobal 缺血的神经保护作用。”

Koji Iihara: "Ischemia induces the expression of PDGF-B chain in neurons..." Journal of Cerebral blood Flow and Metabolism. (発表予定). (1994)

Koji Iihara：“缺血诱导神经元中 PDGF-B 链的表达......”《脑血流与代谢杂志》（即将出版）。