miRNA137/Furin蛋白链对癫痫的修饰及机制研究

The excitatory / inhibitory imbalance in brain play a key role in epileptogenesis. Abnormal synaptic transmission is a key factor which causing changes of excitability. Using a variety of experimental methods, we found that the levels of miRNA137 and Furin proten in regulating synaptic transmission are abnormal regulating in the anterior temporal neocortex of intractable epilepsy patients than control patients in our previous study. This phenomenon shows that miRNA137/Furin chain may involved in epileptogenesis. Under the support of National Natural Science Foundation of China, we have finished the project of: The role and mechanism of miRNA in epilepsy integrin hypotheses. We will continue use our established outpatient clinical resource bank which contains more than 140000 cases of epilepsy patients, brain tissue bank which contains more than 300 cases of brain tissues from intractable epilepsy patients, cerebrospinal fluid bank which contains more than 200 cases of epilepsy patients and several kinds of epilepsy animal models for our research. Using the in vivo multi-channel electrophysiological recording for LFP in hippocampus, the whole cell patch clamp and other molecular biotechnology research techniques, to explore the possible mechanism of miRNA137/Furin chain involved in epileptogenesis through gene over-expression and knock-down in mice epilepsy models and cell epileptic-form model. We aim to explore the role of miRNA137/Furin chain in epileptogenesis by regulating synaptic transmission, to provide a necessary evidence for discovery of a new target for treatment of epilepsy.

脑部兴奋/抑制性失衡在癫痫的形成中起到了核心作用。突触传递异常是引起脑部兴奋性改变的重要因素。课题组前期采用多种实验方法发现与突触传递有关的miRNA137及Furin蛋白在难治性癫痫患者脑组织中具有差异表达，可能参与了癫痫的形成。本课题在已结题的国家自然科学基金项目（编号：81271445）的研究基础上，继续依托课题组已经建立的14000多例癫痫患者门诊临床资料库，300多例颞叶癫痫患者术后脑组织，200多例癫痫患者脑脊液以及多种癫痫动物模型，通过对癫痫模型动物和细胞癫痫样放电模型中的miRNA137/Furin蛋白进行过表达和沉默干预，结合在体多通道电生理记录、全细胞膜片钳记录、生物信息学技术等多种研究手段从表达、功能、机制三个不同层面深入探讨miRNA137/Furin蛋白链在癫痫发作过程中的突触调节作用，揭示其参与调控癫痫发生发展的可能机制，为寻找抗癫痫治疗的新靶点提供理论依据。

1.项目背景： . 本课题是在课题组前期发现与突触传递有关的miRNA137及Furin蛋白在难治性癫痫患者脑组织中具有差异表达基础上，对miRNA137/Furin蛋白链参与调控癫痫发生发展的可能机制进行研究。.2.主要研究内容. 采用癫痫患者术后脑组织、癫痫模型动物、细胞癫痫样放电模型等，结合在体多通道电生理记录、全细胞膜片钳记录、分子生物学技术研究miRNA137/Furin蛋白链对癫痫发生发展的影响。.3.重要结果和关键数据. 1）miRNA137/Furin蛋白在癫痫患者术后脑组织和癫痫模型小鼠脑组织内均有差异表达，且Furin蛋白主要在神经元中表达。. 2）过表达或抑制miRNA137/Furin蛋白链的功能可对癫痫模型小鼠行为学产生明显影响。. 3）过表达或抑制miRNA137/Furin蛋白链的功能可通过调控椎体神经元微小抑制性突触后电流（mIPSCs）来影响神经元的兴奋性。这一结果表明miRNA137/Furin蛋白链参与癫痫的发作可能与GABA介导的抑制性突触传递机制有关。.4.取得的阶段性成果. 研究中先后发表SCI收录论文3篇，揭示了miRNA137/Furin蛋白链参与调控癫痫发生发展的可能机制。