circ-DROSHA通过ceRNA方式调控Ephrin-B3在癫痫突触重塑中的机制研究

研究表明环状circ-DROSHA在癫痫脑组织中表达下调，参与癫痫突触重塑的过程。前期我们研究显示轴突导向分子Ephrin-B3在匹罗卡品癫痫大鼠脑组织中有下调，因此我们提出科学假设circ-DROSHA靶向Ephrin-B3参与突触重塑。本研究拟采用逆转录病毒干预、qPCR、蛋白质共沉淀等技术探讨两者在癫痫大鼠海马神经元突触重塑中的作用；同时应用慢病毒过表达和knockdown circ-DROSHA后，检测Ephrin-B3表达变化，分析两者的正相关性；结合生物信息学的方法筛选与两者有共同结合位点的miRNAs，环状RNA免疫沉淀（circRIP）验证结合力强的miRNAs，通过干预其表达后检测Ephrin-B3表达变化，探索circ-DROSHA通过竞争性内源RNA（ceRNA）方式调控Ephrin-B3在癫痫突触重塑中的作用，为癫痫的治疗提供新的理论依据。

Studies have suggested that circ-DROSHA expressed lower in the temporal lobe epilepsy patients, which play a critical role in the synaptic function maintaining. Our previous studies have showed the axon guidance moleculese ephrin-B3 decreased in epileptic rats. Therefore, we hypothesize circ-DROSHA can modulate Ephrin-B3 in the synaptic remodeling post epilepsy. In this study, we first investigate the expression of circ-DROSHA and ephrin-B3 gene by retrovirus transfection, and then techniques of qPCR, immunoprecipitation would be applied to investigate the role of circ-DROSHA and ephrin-B3 in synaptic remodeling. Furthermore, we detect the expression of ephrin-B3 after circ-DROSHA overexpression and knockdown to clarify the correlation of circ-DROSHA and ephrin-B3. we will clarify the mechanism of circ-DROSHA regulating ephrin-B3 by using biological information analysis and explored a group of miRNAs which might influence ephrin-B3 and circRNA immunoprecipitation （circRIP）was used to verify the miRNAs bind with circ-DROSHA. Then intervene the expression of miRNAs to investigate the influence in ephrin-B3 and find the way of circ-DROSHA regulate Ephrin-B3 by competing endogenous RNAs（ceRNA） in synaptic remodeling in epilepsy, which may provide therapeutic strategies for epilepsy.