调控ROCK和Olig1信号对脑缺血后神经再生作用的研究

项目名称： 调控ROCK和Olig1信号对脑缺血后神经再生作用的研究

项目编号： No.30870855

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 电工技术

项目作者： 张拥波

作者单位： 首都医科大学

项目金额： 28万元

中文摘要： 采用尼龙线栓塞法制备Wistar大鼠大脑中动脉闭塞（MCAO）局灶性脑缺血动物模型，通过Rho激酶抑制剂和腺病毒载体转基因技术来调控ROCK和olig信号，利用病理形态学和伊文思蓝探讨脑缺血后血脑屏障破坏规律；并利用光镜、电镜技术，研究脑缺血后神经元及其线粒体等细胞器的变化；利用免疫组化和免疫印迹法(Western blot)等方法，研究脑缺血后神经血管单元相关蛋白及凋亡相关蛋白、ROCK蛋白质的变化；并观察其对脑缺血后轴突再生和髓鞘修复的影响，探讨了Rho激酶及olig信号对脑缺血后神经元、新生血管、血脑屏障和基质蛋白的作用及其可能机制。结果显示，Rho激酶抑制剂法舒地尔可减轻脑缺血后血脑屏障的破坏，减轻神经细胞的凋亡，保护神经元及其线粒体等细胞器，促进缺血后轴突和髓鞘的再生；腺病毒载体 Ad5-Olig1可促进脑缺血后髓鞘的修复。该研究从整体和全面角度来探讨ROCK对脑缺血后神经血管单元功能调控，同时，结合腺病毒转基因技术探讨Olig1的髓鞘修复作用，从轴突再生和髓鞘修复两个方面，探讨对脑缺血后神经再生的协同效果，为脑缺血治疗提供新的靶点和全面的评价体系。

中文关键词： 脑缺血；大鼠；ROCK信号；Olig1；神经血管单元

英文摘要： The focal cerebral ischemia was made by inducing nylon thread to occlude the middle cerebral artery in Wistar rat. Using Rho kinase inhibitor and adenovirus vector transgene technique regulate the ROCK and olig1 signal pathway. The blood-brain barrier (BBB) disruption was determined by the Evans blue dye extravasations (EBE) technique. The changes of neuron and its mitochondria and other sub-cell organisms were studied using pathology and electron microcopy. The associated proteins with neurovascular unit and apoptosis were studied with immunohistochemistry and Western blot method after ischemia. Axon regeneration and myelin repair were evalued after ischemia. The effects and the mechanisms of Rho kinase and Olig1 signal on the neuron, vessel, BBB and matrix proteins were studied. The results showed that Rho kinase inhibitor Fasudil remarkably ameliorates ischemia-induced BBB disruption, reduces neuronal cell apoptosis, and protect neuron and its mitochondria and other sub-cell organisms, and improve axon regeneration and myelin repair after focal ischemia. Adenovirus vector ad5-Olig1 administrated ventically may improve myelin base protein (MBP) expression in the ipsilateral striatem on 28 days after focal cerebral ischemia in rat. It suggust that Olig1 improve myelin repair after ischemia. This study provide new targets for the treatment of cerebral ischemia from regulating ROCK pathway and olig1 signal, and combinating axon regeneration and myelin repair.

英文关键词： cerebral ischemia; rat; ROCK signal; Olig1; neurovascular unit