异丙酚通过内源性大麻素系统实现心脏保护的机制研究

Perioperative myocardial ischemia reperfusion injury (I/R) contributes to longer hospital stay, more hospitalization cost and higher morbidity and morality. Propofol, an intravenous anesthetic has been found to be effective in reducing myocardial damages caused by ischemia and reperfusion. There are several proposed mechanisms for cardioprotection by propofol: antioxidant-like effect and activation of ROS/NO signaling, inflammation control, and activation of the reperfusion injury salvage kinase (RISK). But how and what is needed for propofol to achieve these effects are unclear. Propofol is a competitive inhibitor fatty acid amide hydrolase (FAAH), which catalyzes the degradation of endocannabinoids (ECs) such as anandamide (AEA) and 2-arachidonoylglycerol (2-AG) . Our previous research has found that propofol post-conditioning can protect against cardiomyocyte apoptosis and death in hypoxia/reoxygenation injury by activating ERK1/2 and JNK signaling, reducing ROS levels and enhancing cell autophagy which are eliminated by pretreatment of selective CB2R antagonist AM630. We therefore proposed a probable role of ECs signaling in cardioprotection by propofol preconditioning and postconditioning. This project deals with the following topics in vivo and in vitro: (1) Regulation of ECs release during propofol preconditioning and postconditioning; (2) Relationship between ECs and cardioprotection by propofol; (3)Roles of CB1R and CB2R signaling in cardioprotection by propofol. The methods for the problems selected include LC/LC-MS method, flow cytometry, Laser Confocal Scanning Microscope imaging, ELISA, Real-Time PCR, Western Blotting, and et al. The goal is to eludicate the function and mechanisms of ECs signaling in propofol-induced cardioprotection in the rat.We hope to publish 1-2 articles cited by SCI and training 1 doctoral student during this research process.

围手术期心肌缺血再灌注损伤发生率高，危害严重，防治困难。动物和临床研究证实，全麻药异丙酚预处理和后处理能够减轻心肌缺血再灌注损伤。既往研究多关注于异丙酚的下游保护性信号通路，对上游激活分子和通路并不清楚。我们前期研究发现，异丙酚后处理可通过激活大麻素CB2受体依赖的ERK1/2、JNK通路，调节ROS生成，增加心肌细胞缺氧/复氧后自噬，减少心肌细胞凋亡和坏死。结合相关文献，我们假设：异丙酚可能通过抑制内源性大麻素（ECs）降解酶FAAH，增加ECs释放，并激活CB1R、CB2R及下游信号，实现心脏保护。拟在细胞和在体水平，采用LC/LC-MS法、免疫荧光和激光共聚焦检测、流式细胞术、ELISA、PCR、Western Blotting等技术手段，从异丙酚调节ECs释放和CB1R、CB2R表达、ECs水平与异丙酚心脏保护的关系、受体机制及信号转导三方面，验证该设想，并发表SCI论文1-2篇。

围术期心肌缺血再灌注损伤发生率高，危害严重，目前尚无有效防治手段，有必要深入研究全麻药预/后处理的心脏保护机制。动物和临床研究证明，全麻药异丙酚预处理和后处理能够减轻心肌缺血再灌注损伤。既往研究多关注于异丙酚的下游保护性信号通路，对上游激活分子和通路并不清楚。我们前期研究发现，异丙酚后处理可通过激活大麻素CB2受体依赖的ERK1/2、JNK通路，调节ROS和NO生成，增加心肌细胞缺氧/复氧后自噬，减少心肌细胞凋亡和坏死。结合相关文献，我们假设：异丙酚可能通过抑制内源性大麻素（ECs）降解酶FAAH，增加ECs释放，并激活CB1R、CB2R及下游信号，实现心脏保护。拟在细胞和在体水平，采用LC/LC-MS法、免疫荧光和流式细胞术、ELISA、western blotting等技术手段，从异丙酚调节ECs释放和CB1R、CB2R表达、ECs水平与异丙酚心脏保护的关系、受体机制及信号转导三方面，验证该设想，为减轻围术期心肌缺血再灌注损伤提供新的思路。