【摘要】 目的 探讨特异性的抑制Calpain能否通过干预Calpain/GSK-3β信号通路降低酒精致大鼠心肌细胞凋亡。方法 100只雄性Wistar大鼠随机分成生理盐水对照组（n=10）、酒精组（n=30）、酒精+Calpain-1抑制剂（ALLN）组（n=30）、酒精+二甲基亚砜溶剂（DMSO）组（n=30）。造模成功后分别通过心脏超声、HE染色、电镜、TUNEL检测、免疫组化染色法及Western blot法检测各组大鼠心肌细胞中Calpain-1、GSK-3β(H-76)、caspase-9 p10、caspase-3 p17的表达情况。结果 与对照组相比，酒精组、酒精+ALLN组、酒精+DMSO组大鼠心脏左室舒张末期内径（LVDD）增大，左室后壁舒张末期厚度（LVPWD）增厚（P<0.05），各组大鼠左室射血分数（LVEF）、左室短轴缩短率（LVFS）均位于正常范围（P>0.05）。酒精组心肌纤维排列紊乱，结构消失，心肌细胞固缩，细胞间质水肿，可见大量炎性细胞浸润，线粒体肿胀，酒精组及酒精+ALLN组心肌细胞凋亡率增加（P<0.05），酒精组Calpain-1、GSK-3β(H-76)、caspase-9 p10、caspase-3 p17蛋白表达水平上调（P<0.05）。酒精+ALLN组与酒精组相比，LVDD略小，LVPWD相对稍薄（P<0.05），病理改变较酒精组减轻，心肌细胞凋亡明显减少（P<0.05）,Calpain-1、GSK-3β(H-76)、caspase-9 p10、caspase-3 p17蛋白表达水平下调（P<0.05）。结论 大量酒精摄入可致大鼠早期酒精性心肌病模型中心肌细胞凋亡水平升高；Calapin-1抑制剂具有降低酒精诱导的心肌细胞凋亡的保护效应，其机制可能与Calpain对GSK-3β的片段化导致其活性增高，上调凋亡相关蛋白的表达有关。更多还原

【Abstract】 Objective To investigate whether or not specific inhibition of Calpain reducing myocardial apoptosis induced by alcohol through interfering Calpain/GSK-3β signaling pathway in rats. Methods Male Wistar rats(n=100) were randomly divided into control group(n=10), alcohol group(n=30), alcohol+Calpain-1group(alcohol+ALLN group, n=30) and alcohol+DMSO group(n=30). After modeling successfully, the expressions of Calpain-1, GSK-3β(H-76), caspase-9 p10 and caspase-3 p17 were detected by using cardiac ultrasound,HE staining, electron microscope, TUNEL detection, immunohistochemical staining(IHC) and Western blotting assay. Results The levels of left ventricular end-diastolic inner diameter(LVDD) and left ventricular posterior wall thickness at end diastole(LVPWD) increased in alcohol group, alcohol+ALLN group and alcohol+DMSO group compared with control group(P<0.05), and left ventricular ejection fraction(LVEF) and left ventricular fraction shortening(LVFS) were normal in all groups(P>0.05). There were disordered myocardial fibers, disappeared structure, cardiomyocyte pyknosis, intercellular substance edema, inflammatory cell invasion and mitochondrial swelling observed in alcohol group. The cardiomyocyte apoptosis rate increased in alcohol group and alcohol+ALLN group(P<0.05), and the protein expressions of Calpain-1, GSK-3β(H-76), caspase-9 p10 and caspase-3 p17were up-regulated(P<0.05) in alcohol group. The levels of LVDD and LVPWD decreased(P<0.05), pathological changes were relieved, cardiomyocyte apoptosis rate decreased significantly(P<0.05), and protein expressions of Calpain-1, GSK-3β(H-76), caspase-9 p10 and caspase-3 p17(P<0.05) were down-regulated in alcohol+ALLN group compared with alcohol group. Conclusion High alcohol intake can increase cardiomyocyte apoptosis in rat model of early alcoholic cardiomyopathy. Calapin-1 inhibitor had a protective effect of reducing alcohol-induced cardiomyocyte apoptosis, and the mechanism may be related to that Calpain can improve GSK-3β activity through GSK-3β fragmentation and up-regulate expressions of apoptosis-related proteins.更多还原