Agrin在脑卒中后运动促进突触再生中的作用及机制研究

项目名称： Agrin在脑卒中后运动促进突触再生中的作用及机制研究

项目编号： No.81460351

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 张彭跃

作者单位： 昆明理工大学

项目金额： 48万元

中文摘要： 脑卒中引起大量神经元和突触损失，因此卒中后突触再生对功能恢复意义重大。我们前期研究表明：康复运动能促进卒中后学习记忆和运动功能恢复、上调BDNF和突触素表达，提示运动能促进突触再生，但对其机制尚不清楚。Agrin是一个对突触形成和维持起关键作用的蛋白，我们推测运动促进突触再生可能是通过运动上调Agrin，激活CREB继而上调BDNF表达，增强突触再生。为验证这一假说，拟首先检测卒中后Agrin、CREB和BDNF表达谱及运动对其影响；其次通过检测Syn1、GAP43、免疫荧光和电镜分析突触再生数量；并通过电生理和行为学评估这些突触的功能。同时建立体外神经元缺血模型，通过添加Agrin及Knock down其表达探明缺血后突触再生是否存在Agrin→CREB→BDNF→突出再生通路。最后通过动物实验验证运动是否通过该通路促进突触再生。通过体内外研究阐明脑卒中后运动训练促进突触再生的分子机制。

中文关键词： 脑卒中；运动训练；突触发生；神经网络；功能恢复

英文摘要： Stroke lead to a significant loss of neurons and synapses, thus the synaptogenesis is critical for recovery of function after stroke. Our previous researches have shown that early exercise after stroke could significantly promoted the recovery of learning and memory, and coordinated locomotor function.The results of our prepare experiment shown that exercise after stroke upregulated the expression of BDNF and synaptophysin in infarct region, which indicated that exercise could promoted the synaptogenesis after stroke. However, the underlying mechanisms are poorly understood.Agrin is a glycoprotein expressed in whole neural system, and is crital for synaptogenesis and stabilization of synapse. Exercise after stroke improve the synaptogenesis, for the underlying mechanisms , we hypothesized that exercise might upregulate the expression of Agrin, active the transcription factor CREB, and then upregualte some protein that improve the synaptogenesis, such as BDNF, promote the recovery of function. To verify the hypothesis,firstly， we test the expression of Agrin after stroke and the effect of exercise on the expression of Agrin，CREB and BDNF；secondly，determine the quantity of synaptogenesis through testing the expression of Syn1 and GAP43；finally，evaluate the function of synaptogenesis through electrophysiology, electron microscope and behavioral methods. Meanwhile, through culturing ischemic neuron by OGD in vitro, we explore the pathway of Agrin→CREB→BDNF→synaptogenesis. Thereafter, we confirm this pathway in vivo through adding recombinant Agrin and antisene-Agrin. Finally, the molecular mechanism for synaptogenesis by early exercise can be demonstrated through these verification research in vivo and vitro.

英文关键词： Stroke;Exercise;Synaptogenesis;Neural network;Functional recovery