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目的探讨阿霉素(ADR)对心肌细胞损伤中钙超载的机制及脂联素的保护作用。方法采用胰蛋白酶消化法原代培养乳鼠心室肌细胞。选用培养72h的单层心肌细胞,实验分为正常对照组、单纯ADR组、ADR+APN(3μg/ml)组、ADR+APN(10μg/ml)组、ADR+APN(20μg/ml)组、ADR+APN(30μg/ml)组。实验终止后,在倒置相差显微镜下观察心肌细胞形态的变化,测定乳酸脱氢酶(LDH)的释放及肌酸激酶(CK)的活性,通过流式细胞术来检测心肌细胞的凋亡,激光共聚焦检测细胞内钙离子荧光强度。结果与对照组相比,给予ADR后,细胞凋亡率显著增加(70.20±3.73vs4.73±1.21,p<0.05),乳酸脱氢酶(LDH)的活性及CK的活性增加(p<0.05),钙离子荧光强度明显增加(605.99±45.62vs137.17±37.7,p<0.05),APN预处理后给予ADR,可较大程度地逆转上述指标变化,与ADR组相比均具有显著性差异(p<0.05),并且呈现一定的浓度依赖性。结论脂联素对阿霉素中毒心肌细胞具有保护作用,其机制可能与保护心肌细胞内质网有关。
Objective To investigate the mechanism of adriamycin (ADR) on calcium overload in cardiomyocytes and the protective effect of adiponectin. Methods Primary cultured neonatal rat ventricular myocytes were isolated by trypsin digestion. The cells were cultured in ADR + APN (10μg / ml) group, ADR + APN (20μg / ml) group, ADR + ADR + APN (30 μg / ml) group. After the termination of the experiment, the morphological changes of myocardial cells were observed under an inverted phase contrast microscope, and the release of lactate dehydrogenase (LDH) and the activity of creatine kinase (CK) were measured. The apoptosis of cardiomyocytes was detected by flow cytometry. Confocal detection of intracellular calcium fluorescence intensity. Results Compared with the control group, the apoptotic rate of ADR was significantly increased (70.20 ± 3.73 vs 4.73 ± 1.21, p <0.05), the activity of lactate dehydrogenase (LDH) and the activity of CK increased (p <0.05 ), The fluorescence intensity of calcium ion increased significantly (605.99 ± 45.62vs137.17 ± 37.7, p <0.05). After pretreatment with APN, the ADR was pretreated, which could reverse the changes of above indexes to a great extent, and had significant difference compared with ADR group (p <0.05), and showed a certain concentration-dependent manner. Conclusion Adiponectin has a protective effect on doxorubicin-induced cardiomyocytes, and its mechanism may be related to the protection of cardiomyocyte endoplasmic reticulum.