c-Abl调控神经元凋亡在癫痫形成中的机制研究

Apoptosis of hippocampal neurons after seizures is an important pathophysiology for refractory epilepsy. c-Abl is a ubiquitous non-receptor tyrosine kinase involved in synaptic plasticity and neuronal apoptosis. Our previous study found that c-Abl expression is increased in patients with epilepsy and a rat model. However, the mechanism of c-Abl involved in epileptogenesis is still to be elucidated. It has been reported that Cdk5 activation by c-Abl leads to neuronal apoptosis. Therefore, we hypothesize that c-Abl regulates neuronal apoptosis in the hippocampus after seizures via Cdk5, leading to increased neuronal excitability and recurrent spontaneous seizures. To verify our hypothesis, we designed a study based on inhibitor or lentivirus-shRNA intervention in pilocarpine-induced chronic spontaneous seizures rat model and Pentylenetetrazole-induced epilepsy model, combined with related techniques in molecular biology, cell biology, behavior and electrophysiology. This project will provide novel insight on the relationship between c-Abl, neuronal apoptosis, and epileptogenesis by examining the mechanism and potential interventions for c-Abl regulation of neuronal apoptosis in the hippocampus in epilepsy, which may provide a new target for the prevention and treatment of epilepsy.

癫痫后神经元凋亡是导致难治性癫痫的一个重要的病理生理基础。c-Abl是一种在突触可塑性和神经元凋亡中起作用的非受体酪氨酸激酶。本项目前期研究发现c-Abl蛋白在癫痫患者及癫痫大鼠模型中表达上调,但c-Abl在癫痫后神经元凋亡中的作用及机制尚不明确。国外多项研究报道，c-Abl激活Cdk5导致神经元凋亡。故我们提出假说：c-Abl通过Cdk5调控神经元凋亡，导致神经元兴奋性增高，自发性癫痫反复发作。为验证假说，本项目拟通过c-Abl抑制剂STI571及慢病毒c-Abl sh RNA对匹罗卡品及戊四氮两种癫痫动物模型干预，结合分子生物学、细胞生物学、行为学、电生理学等研究技术，以“c-Abl-神经元凋亡-癫痫”为新视点，从“功能-机制”两个层次研究c-Abl 在癫痫后神经元凋亡及癫痫形成中的作用及机制，期望为癫痫的防治提供新的药物作用靶点。

癫痫是一种常见的慢性神经系统疾病，当前治疗只能控制癫痫发作，而不能改变癫痫病理生理。我们的前期实验结果发现c-Abl在癫痫患者及癫痫大鼠模型中表达上调，但c-Abl在癫痫中的作用及机制尚不明确。本项目通过匹罗卡品癫痫模型和戊四氮点燃癫痫模型，采用c-Abl抑制剂和慢病毒基因沉默两种干预方式，结合行为学、病理染色、Western Blot 、膜片钳方法进行研究。研究发现：（1）抑制c-Abl降低癫痫发作易感性及发作频率，明确了c-Abl参与癫痫的发生发展（2）抑制c-Abl减少神经元丢失、神经元凋亡、变性，提示c-Abl调控神经元凋亡参与癫痫发生发展；（3）抑制c-Abl降低其下游分子CDK5的活性及神经元兴奋性，进一步阐释了c-Abl在神经元凋亡及癫痫形成中的作用机制。研究结果将为癫痫提出了新的治疗靶点，更新了癫痫发病机制理论，也将为癫痫的防治提供新的思路。