内侧颞叶癫痫的线粒体功能障碍研究及其易感/突变基因分析

本项目针对MTLE患者和动物模型，从线粒体形态和数目、ATP生成率、呼吸链酶复合物活性、钙稳态、膜电位变化及在体患者大脑代谢情况等多个方面测定海马内线粒体功能变化；利用癫痫细胞模型，逐一抑制线粒体各功能后查看神经元放电情况以及各功能间有无相互影响，探讨线粒体各功能在MTLE发生发展中的作用及其相互作用关系。进一步地，利用二代测序技术，检测MTLE患者中有无线粒体相关遗传易感基因，以及MTLE大鼠致痫前后各个时期中有无线粒体遗传物质突变，有望找出新的遗传易感/致病突变基因。本项目从遗传基因水平、蛋白表达水平以及在体功能水平，首次全面探讨线粒体功能障碍在MTLE发病机制中的作用以及线粒体各功能之间的相互作用关系，并开创性地提出遗传易感因素和致痫后的线粒体基因突变参与调控MTLE慢性癫痫形成，为临床难治性MTLE的研究打开新的视角，为避免触发基因突变因素和寻求新的抗癫痫药物研发靶点提供理论依据。

Based on the MTLE patients and rat models, by detecting the mitochondrial morphology and numbers, ATP production rate, the activity of respiratory chain enzyme complexes, calcium homeostasis, membrane potential, brain energy metabolism, the mitochondrial function in hippocampus will be explored globally. Then, by using cell model of epilepsy, neurons’ epileptiform discharge and interactive relationships will be detected after inhibition of single element of mitochondrial function. This research aims to discuss the role of mitochondrial dysfunction in the developing of MTLE and the interactions among the mitochondrial functions. Furthermore, by using generation sequencing technology, we will explore the genetic susceptibility genes in the MTLE patients and the mitochondrial gene mutations in MTLE rat models before and after epileptogenesis, and in the different stages of MTLE. Then, we try to identify new susceptibility/pathogenic genes of MTLE. From the gene level, protein level to function level in vivo, the research will firstly explore the role of mitochondrial dysfunction in the pathogenesis of MTLE comprehensively and the complicated relationships among either mitochondrial function. And we raised a new idea that susceptibility genes and mitochondrial gene mutations involving in the epileptogenesis of MTLE. The study will open new perspectives for the study of clinical refractory MTLE, providing a theoretical basis in order to avoid triggering factors of gene mutation and to identify optimum new biomarkers for pharmacological therapies of MTLE.