线粒体自噬-NLRP3炎症小体途径在脑缺血中的作用及三七总皂苷干预的研究

The finished research manifested that mitophagy had protective effect against cerebral ischemia-reperfusion injury, but the mechanism of this action is unclear. Based on the negative regulatory role of mitophagy in the NLRP3 inflammasome activation and the inflammation plays an important role in cerebral ischemia, we put forward the following hypothesis, cerebral ischemia reperfusion can activate mitophagy, inhibit NLRP3 inflammasome activation, play a protective role in cerebral ischemia reperfusion injury by inhibiting the inflammation. According to this hypothesis, We use the pharmacology, cell and molecular biological techniques to reveal the physiological and pathological significances of mitophay-NLRP3 inflammasome pathway in cerebral ischemia reperfusion injury developed in rat brains in vivo, and in the mouse neurons in vitro. Furthermore, we research the mechanism of panax notoginseng saponins against cerebral ischemia reperfusion injury from the mechanisms of autophagy and elucidate the new mechanism by studying the mitophay-NLRP3 inflammasome pathway. The result of this project will be of benefit to understanding the molecular basis of cerebral ischemia and provide new potential therapeutic targets to treat cerebral ischemia, moreover, this project will further understand the role of PNS in treating cerebral ischemia, provide a theoretical guidance and scientific basis for further development and utilization of the effective ingredients of Panax notoginseng.

已有的研究表明线粒体自噬在脑缺血再灌注损伤中发挥保护作用，但是作用的机制不清楚。基于线粒体自噬抑制NLRP3炎症小体的活化从而抑制炎症反应以及炎症反应在脑缺血中的重要作用，我们提出以下假说，脑缺血再灌注可以激活线粒体自噬，抑制NLRP3炎症小体的活化，通过抑制炎症反应，在脑缺血再灌注损伤中起到重要作用。基于这一假说，我们采用在体和离体脑缺血再灌注模型，利用药理学、细胞生物学和分子生物学技术，揭示线粒体自噬-NLRP3炎症小体途径在脑缺血再灌注损伤中的病理生理学意义，并从自噬调节机制研究三七总皂苷的抗脑缺血再灌注损伤机制，从线粒体自噬-NLRP3炎症小体途径揭示三七总皂苷抗脑缺血再灌注损伤的新机制。本项目的结果将有利于深入了解脑缺血的分子机制，为其治疗提供新的潜在靶点，同时加深对三七总皂苷抗脑缺血作用的认识，为三七有效成份新药的进一步开发和利用提供科学依据。

已有的研究表明NLRP3炎症小体作为炎症反应的核心介导了脑缺血再灌注损伤，而线粒体自噬在脑缺血再灌注损伤中发挥保护作用，并且NLRP3炎症小体的活化受到线粒体自噬的调控，但是在脑缺血再灌注中线粒体自噬是否能够调控NLRP3炎症小体活化，即线粒体自噬-NLRP3炎症小体途径是否介导了脑缺血再灌注损伤及其在脑缺血再灌注损伤中的作用还不明确。本项目综合应用利用药理学、细胞生物学和分子生物学技术，揭示线粒体自噬-NLRP3炎症小体途径在脑缺血再灌注损伤中的病理生理学意义，并从自噬调节机制研究三七总皂苷的抗脑缺血再灌注损伤机制，从线粒体自噬-NLRP3炎症小体途径揭示三七总皂苷抗脑缺血再灌注损伤的新机制。研究结果表明，在脑缺血再灌注中存在这线粒体自噬和NLRP3炎症小体的活化，促进线粒体自噬可以抑制NLRP3炎症小体活化减轻脑缺血再灌注损伤，这表明线粒体自噬-NLRP3炎症小体途径介导了脑缺血再灌注损伤并发挥保护作用；此外，三七总皂苷可以激活线粒体自噬进而抑制NLRP3炎症小体活化减轻脑缺血再灌注损伤，这表明线粒体自噬-NLRP3炎症小体途径是三七总皂苷抗脑缺血再灌注损伤的重要机制。本项目的发现为深入研究缺血性脑损伤的病理生理机制寻找新的干预靶点提供了新的思路，为三七有效成份新药的进一步开发和利用提供了科学依据。