TMEM166基因在小鼠脑缺血再灌注损伤中的功能研究

项目名称： TMEM166基因在小鼠脑缺血再灌注损伤中的功能研究

项目编号： No.81500996

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： 李丽

作者单位： 首都医科大学

项目金额： 19万元

中文摘要： 脑中风是导致全球病人致死和致残的重要原因。我们研究发现TMEM166与脑缺血损伤后的神经元自噬及由此导致的神经损伤密切相关。本研究在TMEM166基因敲除鼠脑缺血再灌注损伤模型(MCAO)的基础上观察其作用并重点探讨其机制。.AMPK-mTOR和 Beclin-1是脑缺血后自噬的重要调节因子。我们的预实验证实在脑缺血阶段AMPK-mTOR被激活，IR阶段AMPK失活Beclin-1升高，TMEM166基因敲除鼠AMPK及 Beclin-1的表达均被抑制。由此我们假想TMEM166分别通过AMPK-mTOR或Beclin-1实现其对MCAO后自噬的调控。本实验从两方面验证假说（1）观察TMEM166诱导的自噬分别在缺血和IR损伤中发挥的作用；（2）采用Compound C、TSC2shRNA、Beclin-1 siRNA及Ad5-TMEM166深入探讨TMEM166作用机制及临床应用前景。

中文关键词： 自噬；缺血再灌注；TMEM166；AMPK-mTOR；Beclin-1

英文摘要： Stroke and its sequelae continue to be a leading cause of disability and mortality worldwide. Despite interventional treatments that significantly decrease the general incidence of stroke, a majority of patients suffer from serious ischemia/reperfusion injury in the process. Our previous study showing TMEM166’s involvement in neuronal excessive autophagy and the subsequent brain damage following ischemia/reperfusion in rats suggests TMEM166 to be a potential novel therapeutic in human stroke. However, the underlying mechanism is unclear and TMEM166’s role in this process requires further cautionary evaluation. In this proposal, we will explore the role of TMEM166 after ischemia/reperfusion brain injury with TMEM166 knock-out mice and focus on the possible neuronal autophagy mechanisms regulated by TMEM166. .Studies have proposed that AMPK-mTOR and Becklin-1 are important regulators of autophagy in response to ischemia/reperfusion injury. Our preliminary results suggest ischemia activates AMPK and inhibits mTOR while reperfusion markedly increases Beclin-1 and rapidly inactivates AMPK. Both AMPK and Beclin-1 expression were inhibited in TMEM166 knock-out animals. Thus, we hypothesized that TMEM166 regulates neuronal autophagy through AMPK-mTOR related mechanisms during ischemia and Beclin-1 during reperfusion. The program contains two primary goals: investigate the role of TMEM166 in a rodent ischemia/reperfusion model and examine the mechanism by which TMEM166 regulates autophagy during ischemia/reperfusion with Compount C (AMPK-specific inhibitor), TSC2shRNA, Beclin-1 siRNA, and adenovirus expression vector (Ad5- TMEM166).

英文关键词： Autophagy; ischemia/reperfusion;TMEM166;AMPK-mTOR;Beclin-1