渥曼青霉素对大鼠缺血再灌注心肌细胞自噬和凋亡的影响  

Effect of wortmannin on autophagy and apoptosis of myocardial cells in rats with ischemia reperfusion

作　　者：王保才[1] 程兆云[1] 葛振伟[1] 

机构地区：[1]河南省人民医院心脏外科,郑州450003

出　　处：《中华实验外科杂志》2016年第2期422-424,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察渥曼青霉素（WT）对大鼠缺血再灌注心肌细胞自噬与凋亡的影响。方法将36只雄性SD大鼠随机分为3组，即假手术组（Sham组，n=12）、心肌缺血再灌注损伤模型组（I／R组，n=12）、I／R＋渥曼青霉素处理组（WT组，n=12）。采用穿线结扎左冠状动脉前降支30min制备I／R模型，WT组于缺血30min后鼠尾静脉注射WT（15μg/kg）。再灌注120min后采集股动脉血样，检测血清乳酸脱氢酶（LDH）及肌酸激酶（CK）活性；取心脏标本，采用氯化三苯基四氮唑染色法（TTC）检测每组大鼠相对心肌梗死面积百分比；取缺血心肌标本，采用原位末端转移酶标记法（TUNEL）检测心肌细胞凋亡，计算细胞凋亡指数（AI），苏木素-伊红（HE）染色法观察心肌细胞病理学变化，Western blot法检测各组心肌细胞中自噬相关蛋白微管相关蛋白1轻链3-Ⅱ（LC3-Ⅱ）及凋亡相关蛋白半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3的表达。结果与I／R组比较，WT组血清LDH[（2356．14±328．46）比（3674．96±409．47）U／LI和CKJ（2412．71±334．84）比（3825．42±433．03）U／L]活性、心肌梗死面积百分比[（39．19±8．34）％比（56．70±9．92）％]及细胞AI[（18．72±3．17）％比（36．58±7．55）％]明显降低（P〈0．01）。WT组心肌细胞损伤较I／R组轻。与I／R组比较，WT组LC3-Ⅱ（0．45±0．18比0．71±0．21）、Caspase-3（0．52±0．23比0．52±0．23）蛋白相对表达量明显降低（P〈0．01）。结论WT干预通过降低心肌细胞自噬活性和细胞凋产水平，减轻大鼠心肌细胞缺血再灌注损伤。Objective To observe the effects of wortmannin （WT） on autophagy and apoptosis in the rat model of myocardial ischemia reperfusion. Methods Thirty - six male SD rats were randomly divided into three groups： sham operation group （Sham group,n = 12）, myocardial ischemia reperfusion injury model group （I/R group,n = 12）, and wortmannin -treated group （WT group, n = 12）. I/R nmdel was prepared by ligation of left anterior descending coronary artery （30 min）. WT （15 g/kg） was injected into the tail vein of rats after ischemia in WT group. 120 rain after reperfusion, femoral artery blood samples were obtained. Serum lactate dehydrogenase （LDH） and creatine kinase （CK） activity was determined. The heart specimens were collected. Triphenyl tetrazolium chloride staining method （TYC） was used to measure the relative myocardial infarction area percentage in each group. . Ischemic myocardial samples were collected, and in situ terminal transferase enzyme labeling method （TUNEL） was applied to detect the apoptosis of myocardial cells, and the apoptosis index （AI） was calculated. The hematoxylin and eosin （HE） staining pathological changes of myocardial cells were observed. By Western blotting, the autophagy related protein microtubule associated protein l light chain 3 （LC3） - lI and apoptosis related cysteinyl aspartatespecific protease （Caspase） -3 protein expression was detected in myocardial cells of each group. Results As compared with I/R group, the activity of serum LDH [ （ 2 356. 14 ± 328.46 ） vs. （3 674. 96±409.47） U/L] and CK [ （2 412. 71 ± 334. 84） vs. （3 825.42 ± 433.03） U/L], the area of myocardial infarction [ （39.19± 8.34） % vs. （56.70± 9.92） % ] and the AI [ （ 18.72 ±3.17 ） % vs. （36. 58± 7.55 ）% ] in WT group were significantly decreased （P 〈 0. 01 ）. The myocardial cell injury in WT group was milder than that in I/R group. As compared with the I/R group, the expression of LC3 - 11 （0. 45±0. 18 vs. 0. 71

关 键 词：渥曼青霉素 心肌缺血 再灌注损伤 白噬 脱噬作用 

分 类 号：R364.12[医药卫生—病理学]