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Molecular mechanisms of neuronal death in ischemia

缺血时神经元死亡的分子机制

基本信息

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

项目摘要

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.

为了阐明缺血性脑损伤神经元死亡的机制，我们观察了沙土鼠前脑缺血后DNA断裂的发生，并评价了几种特异性药物对DNA断裂的影响。通过短暂阻断双侧颈总动脉10分钟造成前脑缺血，并使用TUNEL法证实大脑皮质、尾壳核和海马CA 1区的DNA片段化。环己酰亚胺（蛋白质合成抑制剂）和金精三羧酸（核酸内切酶抑制剂）的管理阻断DNA断裂的发生完全在大脑皮层和部分在尾壳核和海马。GYKI 52466（非NMDA受体抑制剂）和亮抑酶肽（钙蛋白酶抑制剂）给药也观察到类似的作用，但MK-801（NMDA受体抑制剂）给药未显示出阻断作用。为探讨脑缺血相关基因ICE家族在缺血性神经元死亡中的作用，采用北方印迹法检测了大鼠局灶性脑缺血后ICE和YAMA基因的表达。ICE mRNA在缺血后无明显变化，但YAMA mRNA在缺血后10-20小时明显增加，我们正在建立可重复的小鼠缺血模型，以评价凋亡相关基因在缺血性神经元死亡中的作用。