c-Abl在糖尿病大鼠心肌缺血再灌注损伤中的作用  

Role of c-Abl in myocardial ischemia-reperfusion injury in diabetic rats

作　　者：季烨龙 吴洋[1] 冷燕[1] 张艺[1] 张爱宁 夏中元[1] Ji Yelong;Wu Yang;Leng Yan;Zhang Yi;Zhang Aining;Xia Zhongyuan(Department of Anestheiology,Renmin Hospital of Wuhan Universuty,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,430060

出　　处：《中华麻醉学杂志》2020年第5期572-576,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81671891,81970722,81901947)。

摘　　要：目的评价c-Abl在糖尿病大鼠心肌缺血再灌注损伤中的作用。方法SPF级健康雄性SD大鼠60只,7~9周龄,体重210~230 g,采用随机数字表法将大鼠分成6组:假手术组(S组,n=6)、心肌缺血再灌注组(IR组,n=12)、糖尿病假手术组(DS组,n=6)、糖尿病心肌缺血再灌注组(DIR组,n=12)、糖尿病心肌缺血再灌注+AVV9-siRNA-c-Abl组(DIR+c-Abl组,n=12)和糖尿病心肌缺血再灌注+AVV9-GFP组(DIR+GFP组,n=12)。采用腹腔注射1%链脲佐菌素-柠檬酸盐缓冲液60 mg/kg的方法制备1型糖尿病模型。DIR+c-Abl组和DIR+GFP组于糖尿病建模后4周尾静脉缓慢注射AVV9-siRNA-c-Abl和AVV9-GFP 1×1012 mg/kg。糖尿病建模后8周,采用结扎冠状动脉左前降支30 min再灌注2 h的方法建立心肌缺血再灌注损伤模型。再灌注结束时记录左心室收缩压(LVSP)和左心室收缩压上升和下降最大速率(±dp/dtmax),取血样,采用ELISA法检测血清LDH和CK-MB浓度,TTC法检测心肌梗死面积(除S组和DS组),TUNEL染色法检测心肌细胞凋亡指数,Western blot法检测心肌组织c-Abl、p53、活化caspase-3的表达水平,免疫共沉淀法检测心肌组织c-Abl与p53的结合水平(c-Abl/p53)。结果IR组与S组相比、DIR组与DS组相比LVSP和±dp/dtmax降低,血清LDH和CK-MB浓度升高,凋亡指数和c-Abl/p53升高,c-Abl、p53和活化caspase-3表达上调(P<0.05);与DIR组相比,DIR+c-Abl组LVSP和±dp/dtmax升高,血清LDH和CK-MB浓度降低,心肌梗死面积减小,凋亡指数和c-Abl/p53降低,c-Abl、p53和活化caspase-3表达下调(P<0.05),DIR+GFP组上述指标差异无统计学意义(P>0.05)。结论c-Abl可能通过激活p53信号通路,参与糖尿病大鼠心肌缺血再灌注损伤的病理生理过程。Objective To evaluate the role of c-Abl in myocardial ischemia-reperfusion(I/R)injury in diabetic rats.Methods Sixty male SPF-grade healthy Sprague-Dawley rats,aged 7-9 weeks,weighing 210-230 g,were divided into 6 groups by a random number table method:sham operation group(S group,n=6),myocardial I/R group(IR group,n=12),diabetic sham operation group(DS group,n=6),diabetic myocardial I/R group(DIR group,n=12),diabetic myocardial I/R plus AVV9-siRNA-c-Abl group(DIR+c-Abl group,n=12),and diabetic myocardial I/R plus AVV9-GFP group(DIR+GFP group,n=12).One percent streptozotocin 60 mg/kg was intraperitoneally injected to induce type 1 diabetes mellitus.AVV9-siRNA-c-Abl and AVV9-GFP 1×1012 mg/kg were slowly injected at 4 weeks after establishing the model in DIR+c-Abl and DIR+GFP groups.Myocardial ischemia was induced by 30 min occlusion of left anterior descending branch(LAD)of coronary artery followed by 2 h reperfusion at 8 weeks after establishing the model.At the end of reperfusion,left ventricular systolic pressure(LVSP)and the maximum rate of increase and decrease of left ventricular systolic pressure(±dp/dtmax)were monitored,and blood samples were collected for determination of the concentrations of serum lactic dehydrogenase(LDH)and creatine kinase-MB(CK-MB)(by enzyme-linked immunosorbent assay)and myocardial infarct size(except group S and group DS).Myocardial tissues were obtained for determination of the apoptosis index of cardiomyocytes(by TUNEL staining),expression of c-Abl,p53 and activated caspase-3(by Western blot),and binding of c-Abl and p53(c-Abl/p53)(by co-immunoprecipitation method).Results LVSP and±dp/dtmax were significantly decreased,serum LDH and CK-MB concentrations were increased,apoptosis index and c-Abl/p53 were increased,and the expression of c-Abl,p53 and activated caspase-3 was up-regulated in group IR when compared with group S and in group DIR as compared with group DS(P<0.05).Compared with group DIR,the LVSP and±dp/dtmax were significantly increased,serum LDH and CK-MB concentra

关 键 词：原癌基因蛋白质c-abl 糖尿病 心肌再灌注损伤 

分 类 号：R-332[医药卫生] R587.1R619