基于NLRP3炎症小体/IL-1β/CCL2-CCR2介导血脑屏障破坏探讨柴贝止痫汤治疗耐药性癫痫的作用机制

耐药性癫痫（DRE）严重危害人类健康，缺乏有效治疗手段。神经炎症及其介导的血脑屏障（BBB）损伤是DRE主要病理特征。NLRP3炎症小体产生IL-1β进而特异性介导CCL2-CCR2轴激活，放大神经炎症反应破坏BBB结构和功能，是导致DRE的关键机制。前期工作已发现柴贝止痫汤可降低多药耐药蛋白P-gp过表达改善BBB转运功能控制DRE发作，但具体机制尚不明晰。基于此申请人提出假说，该复方可能通过NLRP3炎症小体/IL-1β/CCL2-CCR2途径调控脑内免疫微环境，修复BBB结构和功能治疗DRE。本研究拟采用荧光定量PCR、免疫印迹、免疫荧光、透射电镜、罗丹明123实验等方法，通过体内外实验对比观察柴贝止痫汤对NLRP3炎症小体、IL-1β、CCL2、CCR2表达，BBB紧密连接蛋白、药物和谷氨酸转运蛋白，癫痫敏感性的影响，揭示该复方治疗DRE的效用机制，为其进一步临床推广提供依据。

Drug-resistant epilepsy (DRE) seriously harms human health and lacks effective treatment. Neuroinflammation and its mediated blood-brain barrier (BBB) damage are the main pathological features of DRE. NLRP3 inflammasome produces IL-1β and then IL-1β specifically mediates the activation of the CCL2-CCR2 axis, amplifies the neuroinflammatory response and destroys the structure and function of the BBB, which is the key mechanism leading to DRE. Previous work has found that Chaibei Zhixian Decoction can reduce the overexpression of the multidrug resistance protein P-gp and improve the BBB transport function to control the seizure of DRE, but the specific mechanism is still unclear. The hypothesis proposed by the applicant based on the previous work: Chaibei Zhixian Decoction may regulate the immune microenvironment in the brain through the NLRP3 inflammasome/IL-1β/CCL2-CCR2 path, so as to repair the structure and function of the BBB and treat DRE. In this study, RT-PCR, Western Blotting, Immunofluorescence, Transmission Electron Microscopy, Rhodamine 123 Experiment were used to compare the influence of Chaibei Zhixian Decoction on NLRP3 inflammasome, IL-1β, CCL2, CCR2, BBB tight junction protein, drugs and glutamate transporters, epilepsy sensitivity by the experiments in vitro and in vivo. We hope to reveal the effective mechanism of Chaibei Zhixian Decoction in the treatment of DRE, and provide a basis for its further clinical promotion.