长链非编码RNA H19在颞叶癫痫海马神经元凋亡中的作用及调控机制

项目名称： 长链非编码RNA H19在颞叶癫痫海马神经元凋亡中的作用及调控机制

项目编号： No.81471315

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 孟凡刚

作者单位： 北京市神经外科研究所

项目金额： 70万元

中文摘要： 癫痫是常见的严重威胁人类健康的神经系统慢性疾病，其病因复杂且不明确。前期工作中，我们对颞叶癫痫大鼠海马组织长链非编码RNA（lncRNA）表达谱进行了筛查、生物信息学分析及PCR验证，发现癫痫大鼠海马组织lncRNA发生显著性差异改变，其中lncRNA H19上调达8倍以上，而且提示H19与凋亡具有强相关性。鉴于细胞凋亡在癫痫中的作用，我们推测lncRNA H19在颞叶癫痫所致的海马神经元凋亡中有重要的调控作用。本课题中，申请者拟分别从在体、离体层面，通过构建干扰慢病毒载体、细胞转染、RIP-seq、Pull-down、PCR、免疫组化及Western-Blot等方法从分子、细胞及整体水平系统深入地研究H19的功能及作用方式，阐明H19在颞叶癫痫海马神经元凋亡中的作用及调控机制，为研究癫痫发病机制提供理论基础，并为颞叶癫痫疾病提供新的治疗靶点。

中文关键词： 颞叶癫痫；发病机制；长链非编码RNA；H19；细胞凋亡

英文摘要： Epilepsy is a common chronic disease of the nervous system threating to human health seriouslly, of which the etiology is complex and not very clear. In previous work, we screened the lncRNA expression profile of the hippocampus of the temporal lobe epilepsy (TLE), and validated through PCR. The most up-regulated lncRNA in hippucampal tissues was H19 with more than 8 fold, which prompted the strong correlation between H19 and apoptosis. In view of the role that cell apoptosis played in epilepsy, these observations led to our hypothesis that,H19 may play an important role in the development of epilepsy. In our current project, we intend to carry out in vivo and in vitro experiments,through the methods of constructing interference lentiviral vectors, RIP-seq, RNA Pull-down, PCR, immunohistochemistry and western blot, to investigate the lncRNA function and mechanism of action from the levels of the molecular, the cellular and the integral. So as to clarify the function and the regulatory mechanism, by which H19 played in the apoptosis in hippocampal neurons of the TLE, which is to provide the theoretical basis for the mechanism and the therapeutic targets of epilepsy.

英文关键词： epilepsy;etiology;RNA; Long Noncoding;H19;Apoptosis