RIP1/RIP3/Drp1信号通路调控蛛网膜下腔出血后NLRP3炎症小体激活的作用研究

蛛网膜下腔出血（SAH）是一种常见的神经急危重症，缺乏有效的药物。SAH后急性炎症反应是脑损伤的重要病理机制之一。我们前期研究证实小胶质细胞NLRP3炎症小体促进SAH后急性炎症；NLRP3小干扰RNA缓解SAH后炎症反应；线粒体ROS为NLRP3炎症小体的激活提供了重要的信号。但SAH后线粒体ROS的调控因素尚不清楚。本项目预实验发现SAH后RIP1的表达明显增强，拟从多个分子水平验证假说：RIP1/RIP3/Drp1信号通路增加线粒体ROS含量从而激活NLRP3炎症小体，促进SAH后急性炎症反应。我们在前期研究工作的基础上，建立SAH大鼠模型和小胶质细胞体外模型，以特异性药物阻断剂和小干扰RNA基因敲除作为干预手段，应用免疫共沉淀，免疫探针及电镜等分子生物学方法，探索SAH后NLRP3炎症小体的激活是否依赖RIP1/RIP3/Drp1信号通路调控，为减轻SAH早期脑损伤提供新策略。

Subarachnoid hemorrhage (SAH) is a severe neurological disorder, which is lack of effective drug. Acute neuroinflammation is one of the pathophysiological mechanisms of SAH-induced brain injury. Our previous study demonstrated that NLRP3 inflammasome in microglia plays an important role in inflammation after SAH. NLRP3 small interfering RNA can ameliorate neuroinflammation after SAH. Mitochondrial ROS can active NLRP3 inflammasome. We hypothesize that RIP1/RIP3/Drp1 pathway takes part in the production of mitochondrial ROS, which subsequently leads to activation of NLRP3 inflammasome after SAH. Our preliminary test observed the increasement of RIP1 after SAH. Based on preliminary data, we establish vascular puncture SAH model in rats, apply specific pharmacological antagonist and small interfering RNA gene of RIP1-RIP3 and Drp1 and explore that RIP1/RIP3/Drp1 signal pathway is involved in regulating NLRP3 inflammasome by immunoprecipitation, immune-probe, electron microscopy, etc. We hope to find a new target for the intervention of acute inflammatory response after SAH.