电针调控MDM2泛素化PSD-95水平重塑海马神经元突触结构改善CFS认知功能障碍的机制研究

海马神经元突触结构受损是慢性疲劳综合征（CFS）认知功能障碍发生的关键机制，我们前期证实电针可以抗海马区炎症损伤，改善CFS模型大鼠学习记忆能力，通过磷酸化蛋白质组学筛选海马中的差异蛋白发现电针干预后与突触结构、功能高度相关的PSD-95显著回调，提示其可能为电针调控的潜在靶点。PSD-95表达水平受泛素-蛋白酶体系统（UPS）调节，MDM2是泛素连接酶E3在海马中的主要表达形式，MDM2对PSD-95异常泛素化与突触结构受损密切相关。因此提出电针可能通过抑制MDM2活性，调整PSD-95的异常泛素化，改善海马神经元突触可塑性的研究假说。本项目以UPS为切入点，运用基因敲除、透射电镜、WB、PCR等技术，结合micro PET/CT观察海马突触重塑脑代谢响应特征，证明PSD-95为电针治疗CFS认知功能障碍的关键效应蛋白。研究结果将从功能评价、形态结构和细胞分子信号三方面解析电针作用机制。

The damage of synaptic structure of hippocampal neurons is the key mechanism of cognitive dysfunction in chronic fatigue syndrome (CFS). We have previously confirmed that electroacupuncture can prevent inflammatory damage in the hippocampal region and improve the learning and memory ability of CFS model rats. The phosphorylation proteomics was used to screen the differential proteins in the hippocampus and found that the PSD-95 significantly correlated with synaptic structure and function after electroacupuncture intervention, suggesting that it might be a potential target for electroacupuncture regulation. The expression level of PSD-95 is regulated by the ubiquitin proteasome system (UPS). MDM2 is the main expression form of ubiquitin ligase E3 in the hippocampus. The abnormal ubiquitination of PSD-95 by MDM2 is closely related to the damage of synaptic structure. Therefore, it is suggested that electroacupuncture may regulate the abnormal ubiquitination of PSD-95 and improve the synaptic plasticity of hippocampal neurons by inhibiting the activity of MDM2. This project will take UPS as the entry point, use gene knockout, transmission electron microscopy, WB, PCR and other technologies, combined with micro PET/CT to observe the characteristics of cerebral metabolic response of hippocampal synaptic remodeling, and prove that PSD-95 is a key effector protein in the treatment of CFS cognitive dysfunction by electroacupuncture. The results will analyze the mechanism of electroacupuncture from three aspects: functional evaluation, morphological structure and cell molecular signal.