EphB4受体对脑卒中后神经干细胞增殖、分化的作用及机制研究

项目名称： EphB4受体对脑卒中后神经干细胞增殖、分化的作用及机制研究

项目编号： No.81503049

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 孙芳玲

作者单位： 首都医科大学

项目金额： 17.9万元

中文摘要： 脑卒中是危害人类健康的第三大疾病，其高致残率和致死率归因于神经元死亡导致的永久性神经功能缺陷。而神经再生受复杂的网络调控，发现促进神经再生修复的新靶点和新方法，是目前脑卒中神经康复治疗的难点。.本研究从细胞和动物2个水平研究EphB4对神经干细胞增殖、分化的作用和分子机制：①构建人胚胎神经干细胞EphB4基因沉默和过表达模型，BrdU、GFAP、TUNEL等荧光染色观察EphB4对神经干细胞增殖、分化、凋亡的影响；使用蛋白抑制剂探讨EphB4与下游Abl、CyclinD1的关系；②构建EphB4基因敲除小鼠，给予脑缺血或电刺激，观察EphB4对神经干细胞增殖、新生细胞成熟以及树突棘密度等突触可塑性的影响，并通过行为学评分和核磁共振成像等观察EphB4基因敲除对脑卒中后神经功能和梗死体积的作用。.本研究将发现一个新的神经干细胞调节机制，为神经再生治疗脑卒中提供新靶点。

中文关键词： EphB4受体；神经干细胞；基因敲除；脑卒中；电刺激

英文摘要： Stroke is a leading cause of disability and mortality worldwide. It is due to neuronal death and thereby irreversed neurological dysfunction after cerebral ischemia. Since neural stem cells renewal is regulated by a complex network, how to find a new beneficial way to promote neurogenesis is thought to be the difficulty in the development of neurorestorative therapy for stroke..The aim of this study is to demonstrate the role of EphB4 receptor in the neural stem cells proliferation and differentiation, in vitro and in vivo. ① Construct EphB4 overexpression model or silence EphB4 expression with siRNA on human embryonic neural stem cells, and then observe the impact on the cell proliferation, differentiation and apoptosis by immunofluoresence detection with BrdU, GFAP, TUNEL and so on. To indicate possible mechanism of EphB4 in regulating cell proliferation, the relationship between EphB4 and downstream proteins Abl, CyclinD1 will be discussed by using protein inhibitor. ② Establish an EphB4 gene knock-out animal model, and determine that if EphB4 is associated with neurogenesis induced by cerebral ischemia or electroconvulsive stimulation, as well as the maturation of new born cells and synaptic plasticity. Effects of EphB4 knock-out on neurological function and infarct volume after ischemic stroke will also be clarified..EphB4 may emerge as a new signaling molecule for regulating the performance of neural stem cell, and this study will provide a novel and promising candidate approach for the neurorestoration treatment of stroke.

英文关键词： EphB4 receptor;neural stem cell ;gene knock-out;stroke;electroconvulsive stimulation