基于Pannexin1/P2X7R-NLRP3炎性小体活化途径研究愈痫灵颗粒治疗癫痫的神经免疫调节作用

癫痫是一种常见神经系统疾病，其发病机制尚未完全明确。目前癫痫的免疫机制成为研究热点，神经免疫网络失衡是其发病的重要因素。研究发现缝隙连接蛋白Pannexin1介导NLRP3炎症小体活化在癫痫在固有免疫及炎症反应的过程中扮演重要的角色。课题组前期研究证实，愈痫灵颗粒可抑制Pannexin1/P2X7R通道及下游的炎性因子的表达。在此基础上，本研究以Pannexin1介导NLRP3炎症小体活化和炎性反应机制为切入点，通过建立氯化锂-匹罗卡品点燃大鼠癫痫模型和无镁细胞外液诱导的细胞模型，从体内、外两条途径，采用多层次(整体，器官，细胞，分子)和多手段(膜片钳，分子生物学，流式细胞仪、电镜等)相结合的方法，研究愈痫灵颗粒对Pannexin1/P2X7R-NLRP3炎性小体活化相关元件调控效应及神经元保护作用，为中药调控神经-免疫网络机制提供分子生物学基础。

Epilepsy is a common nervous system disease, its pathogenesis has not been completely clear. At present, the immune mechanism of epilepsy has become a research focus. The imbalance of neural immune network is an important factor in the pathogenesis of epilepsy. It was found that the activation of NLRP3 inflammatory bodies mediated by Pannexin 1 plays an important role in the innate immune and inflammatory response of epilepsy. Our previous study confirmed that Yuxianling granule can inhibit the expression of pannexin1/P2X7R channel and downstream inflammatory factors. On this basis, this study takes the mechanism of Pannexin 1-mediated NLRP3 inflammatory body activation and inflammatory response as the entry point, through the establishment of lithium pilocarpine kindling rat epilepsy model and magnesium free extracellular fluid induced cell model, from in vivo and in vitro, using multi-level (whole, organ, cell, molecule) and multi-means (patch clamp, molecular biology, flow cytometry Electron microscopy and other methods were used to study the regulatory effect of Yuxianling Granule on the elements related to Pannexin 1 /P2X7R-NLRP3 inflammatory corpuscle activation and the neuron protection, so as to provide molecular biological basis for the regulation of neural immune network mechanism by traditional Chinese medicine.