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目的观察白藜芦醇(resveratrol,Res)对异丙肾上腺素(isoproterenol,ISO)诱导的大鼠心肌细胞凋亡的影响及其机制。方法利用ISO建立乳鼠心肌细胞凋亡模型,分为正常对照组(无血清DMEM培养液)、模型组(无血清DMEM培养液加入ISO 1μmol/L作用48 h),白藜芦醇干预组(分别向培养基中加入ISO 1μmol/L和Res 50μmol/L作用48 h)和白藜芦醇对照组(向无血清DMEM培养液中加入Res 50μmol/L)。Hochest33258染色检测细胞凋亡率;电镜观察心肌细胞超微结构;检测培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)漏出量;Western blot检测Bcl-2和Bax蛋白表达。结果模型组心肌细胞数量减少,细胞核形态不规则,可见凋亡小体,细胞凋亡率增加。白藜芦醇干预组和白藜芦醇对照组细胞膜完整,核膜清晰,核内染色质浓缩,细胞损伤程度减轻。与正常对照组比较,模型组心肌细胞凋亡率和LDH漏出量均增加,Bcl-2蛋白水平下调,Bax蛋白水平上调,差异均有统计学意义(P<0.05,P<0.01);与模型组比较,白藜芦醇干预组和白藜芦醇对照组心肌细胞凋亡率和LDH漏出量均降低,Bcl-2蛋白表达上调,Bax蛋白表达下调,差异均有统计学意义(P<0.05)。结论 Res明显减轻ISO诱导的心肌细胞凋亡,其机制可能与逆转凋亡相关因子Bcl-2和Bax蛋白表达有关。
Objective To investigate the effect of resveratrol (Res) on rat cardiomyocyte apoptosis induced by isoproterenol (ISO) and its mechanism. Methods The apoptosis model of cardiomyocytes was established by ISO and divided into normal control group (serum-free DMEM medium), model group (serum-free DMEM medium supplemented with ISO 1μmol / L for 48 h), resveratrol intervention group Respectively add ISO 1μmol / L and Res 50μmol / L for 48 h) and resveratrol control group (add Res 50μmol / L to serum-free DMEM). Hoechst 33258 staining was used to detect the apoptosis rate. Electron microscopy was used to observe the ultrastructure of myocardial cells. The amount of lactate dehydrogenase (LDH) leakage in the culture medium was detected. The protein expressions of Bcl-2 and Bax were detected by Western blot. Results The number of myocardial cells in the model group decreased, and the nuclear morphology was irregular. The apoptotic bodies were seen and the apoptosis rate increased. Resveratrol intervention group and resveratrol control group complete cell membrane, clear nuclear membrane, chromatin condensation in the nucleus, reduce the degree of cell damage. Compared with the normal control group, the apoptotic rate of myocardial cells and the leakage of LDH in model group were increased, while the levels of Bcl-2 protein and Bax protein were up-regulated in the model group (P <0.05, P <0.01) Compared with the resveratrol group and the resveratrol group, the apoptotic rate of myocardial cells and LDH leakage were decreased, the expression of Bcl-2 protein and Bax protein were down-regulated in the resveratrol group and the resveratrol group (P <0.05) ). Conclusion Res significantly attenuates ISO-induced cardiomyocyte apoptosis, which may be related to reversing the expression of Bcl-2 and Bax protein.