自噬小体清除在七氟烷预处理延迟性心肌保护中的作用  

Role of autophagosomes clearance in delayed cardioprotection in rats with ischemia-reperfusion injury by sevoflurane preconditioning in vivo

作　　者：乔世刚[1,2,3] 孙波[3] 单海华[3] 王安[3] 仇佳[3] 王琛[1,2,3] 

机构地区：[1]苏州科技城医院麻醉科,苏州215153 [2]南京医科大学附属苏州医院麻醉科,苏州215153 [3]苏州大学附属第二医院麻醉科,苏州215004

出　　处：《中华急诊医学杂志》2017年第1期65-70,共6页Chinese Journal of Emergency Medicine

基　　金：江苏省自然科学基金面上研究项目（BK20141187）;苏州市科技计划项目（SYS201473）;苏州大学附属第二医院博士、留学归国人员预研项目（SDFEYBS1401）

摘　　要：目的探讨自噬小体清除水平在七氟烷预处理产生延迟性心肌保护中的作用。方法成年雄性SD大鼠45只，体质量270-350g，采用随机数字表法，随机分为3组（n=15）：假手术组（sham组）；缺血一再灌注组（CON组）；七氟烷延迟性组（SWOP组）。采用结扎左冠状动脉前降支30min后再灌注2h的方法制备在体大鼠心肌缺血-再灌注模型，于再灌注2h时取心肌组织，TYC染色法测定心肌梗死范围；TUNEL试剂盒检测细胞凋亡；透射电镜观察心肌自噬小体变化；Western blot法检测自噬溶酶体相关蛋白LC3-Ⅱ、Cathepsin B、自噬性底物p62以及cleavedcaspase-3的表达水平。采用SPSS15．0统计软件进行数据分析。计量资料以均数±标准差（x±s）表示，组间比较采用单因素方差分析。结果与sham组比较，CON组的心肌凋亡指数增高（P=0．004）、自噬小体增多，LC3-Ⅱ（P=0．009）、Cathepsin B（JP=0．032）、p62（P=0．007）和caspase-3（P=0。006）水平明显升高；与CON组比较，SWOP组的梗死范围（P=0．027）、凋亡指数（P=0．042）、自噬小体减少，LC3-Ⅱ（P=0．033）、p62（P=0．041）和cleaved caspase-3（P=0．037）水平降低，Cathepsin B（P=0．046）水平升高。结论七氟烷预处理对在体大鼠心肌缺血-再灌注产生延迟性保护作用可能与增强心肌自噬小体清除有关。Objective To evaluate role of autophagosomes clearance in delayed cardioprotection by sevoflurane preconditioning in rats with ischemia-reperfusion injury in vivo. Methods Forty-five adult male Sprague-Dawley rats, weighing 270-350 g, were randomly （random number） divided into 3 groups： sham operation group （sham group）, isehemia-reperfusion group （CON group）, sevoflurane preconditioning group （SWOP group）. Myocardial ischemia was induced by 30 min occlusion of left anterior descending branch （LAD） of coronary artery followed by reperfusion for 2 h, and myocardial infarct size was stained by triphenyhetrazolium chloride. Cardiomyoeyte apoptosis was evaluated by terminal deoxyribonucleotidyl transferase-mediated biotin-16dUTP nick-end labeling. Autophagosomes were detected under transmission electron microscope. Expression of LC3-11 , eathepsin B, p62 and cleaved caspase-3 were assessed by western blotting. Statistical analysis were performed using one or two way analysis of variance （SPSS 15.0, Chicago, USA） test followed by Dunnet-t or LSD-t test. Results Sevoflurane preconditioning reduced myocardial infarct size and the number of autophagosomes （ P = 0. 027 ）, attenuated cardiomyocyte apoptosis （P = 0. 042）. Sevoflurane preconditioning decreased the level of LC3-1/ （P = 0. 033 ）, p62 （ P = 0. 041 ） and cleaved caspase-3 （P = 0. 037）, but increased the level of cathepsin B （P = 0. 046）. Conclusions Delayed cardioprotection by sevoflurane preconditioning increased myocardial clearance of autophagosomes against the delayed ischemia reperfusion injury.

关 键 词：麻醉药 吸入 心肌再灌注损伤 自噬 细胞凋亡 溶酶体 

分 类 号：R614[医药卫生—麻醉学]