NLRP3介导的小胶质细胞焦亡在蛛网膜下腔出血中的作用及与LncRNA H19相关的调控机制研究

目前，蛛网膜下腔出血（SAH）尤其是其早期脑损伤的病理机制不明确，临床上也缺乏有效的药物。我们前期研究证实SAH后早期小胶质细胞上NLRP3炎症小体激活是神经炎症发生的关键，但临床研究仅针对其下游炎症因子不能有效改善脑损伤，提示NLRP3炎症小体可能存在其他引起神经损伤的病理机制。预实验发现SAH早期，焦亡关键蛋白Gasdermin D及其活性部分在小胶质细胞表达上升，提示小胶质细胞可能发生NLRP3介导的焦亡；进一步，生物信息学分析和预实验提示NLRP3还可能受到lncRNA H19/miR-138的调控。因此，本研究拟通过构建小胶质细胞特异性NLRP3过表达小鼠和慢病毒介导的过表达或敲减，并应用MRI，荧光素酶基因报告等手段，明确NLRP3介导的小胶质细胞焦亡在SAH早期脑损伤中的作用，以及通过lncRNA H19调控NLRP3表达的机制，以期为解析SAH发病机制及治疗提供新思路。

So far, the pathological mechanism of subarachnoid hemorrhage (SAH), especially early brain injury, remains unclear. Furthermore, there is currently no effective treatment for SAH. Our previous studies confirmed that the activation of NLRP3 inflammasome in microglia plays a key role in inflammatory response early after SAH, but antiinflammation targeting its downstream cytokine alone can not effectively ameliorate brain damage, which suggested there are another other pathological mechanisms in NLRP3 inflammasome-induced brain injury. The preliminary experiment demonstrated that the expression of the key pyroptosis effector protein, Gasdermin D, and its active domain (N-terminal) increased in microglia in the early stage of SAH, which indicated that NLRP3-mediated pyroptosis may occur in microglia. Further, bioinformatics analysis and preliminary experiments suggested the expression of NLRP3 may be regulated by lncRNA H19/miR-138. Therefore, this study aims to explore the role of NLRP3-mediated pyroptosis in microglia in SAH-induced brain injury and lncRNA H19 related regulatory mechanism of NLRP3 by constructing microglia-specific overexpression of NLRP3 mice and lentivirus-mediated overexpression or knockdown, MRI and luciferase reporter gene assays, etc. The study is expected to provide new ideas for the pathogenesis and treatment of SAH.