Drp1调控的线粒体自噬途径在癫痫神经损伤中的作用及机制研究

项目名称： Drp1调控的线粒体自噬途径在癫痫神经损伤中的作用及机制研究

项目编号： No.81201012

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 神经系统疾病、精神疾病

项目作者： 谢南昌

作者单位： 郑州大学

项目金额： 23万元

中文摘要： 癫痫发作后线粒体受损导致神经元对痫性损伤更敏感，及时清除受损线粒体对神经元存活是至关重要的。适当的线粒体自噬可以选择性清除受损线粒体以维持细胞内环境稳态，但线粒体自噬在癫痫神经损伤中的作用及机制目前并不清楚。最近研究发现线粒体融合分裂与线粒体自噬有密切关系，我们前期研究也发现癫痫大鼠海马线粒体分裂蛋白Drp1和线粒体自噬水平均明显升高，因此推测Drp1可能通过对线粒体自噬的调控进而影响癫痫的发生发展。为了证明这一假说，我们将利用体外培养海马神经元癫痫放电及PILO诱导的大鼠颞叶癫痫模型，构建腺病毒载体干预Drp1表达，采用分子生物学、透射电镜及激光共聚焦等技术观察对线粒体自噬、线粒体损伤和细胞凋亡的影响。再给予调控自噬（自噬诱导剂或抑制剂）和线粒体通透转运（开放或抑制mPTP）干预，进一步证明Drp1发挥作用的信号级联。本课题将从新的视觉阐明癫痫神经损伤的分子机制，为癫痫治疗提供新的靶点。

中文关键词： 癫痫；动力相关蛋白1；凋亡；线粒体自噬；内质网应激

英文摘要： Seizure-induced mitochondrial damage makes epileptic neurons more sensitive to injury. Therefore, Timely removal of damaged mitochondria is important for neuronal survival in epilepsy. Appropriate mitophagy can selectively remove damaged mitochondria to maintain the steady-state intracellular environment. However, the function and mechanism of mitophagy in seizure-induced neuronal injury is still unknown. Recent studies have revealed that mitochondrial fusion and fission is closely with mitophagy. Our previous study also found that the expression of the mitochondrial fission protein Drp1（Dynamin-related protein 1，Drp1）and mitophagy were both significantly increased in hippocampus of epileptic rats. So we speculate Drp1 may affect the development of epilepsy by regulation of mitophagy. Using in vitro primary cultured hippocampal neuron epileptic discharge and in vivo PILO（pilocarpine, PILO）-induced rat model of temporal lobe epilepsy, We will detect the effect of Drp1 on mitophagy, mitochondrial injury and apoptosis by molecular biology, transmission electron microscope, and Laser scanning confocal technology by constructing adnoviral vectors to regulate the expression of Drp1. In addition, we will further prove the signaling cascade of Drp1 by regulation of autophagy (induction or inhibition of autophagy) and mi

英文关键词： Epilepsy；Drp1；Apoptosis；Mitophagy；ERS