薯蓣皂苷对缺氧/复氧所致乳鼠心肌细胞凋亡的保护作用

The protective effect of dioscin on cardiomyocytes apoptosis after anoxia/reoxygenation injury

目的:研究薯蓣皂苷对缺氧/复氧所致乳鼠心肌细胞凋亡的保护作用,并探讨其作用机制。方法:体外原代培养乳鼠心肌细胞,建立缺氧/复氧损伤模型,以不同剂量的薯蓣皂苷进行干预。实验分为5组:空白对照组、缺氧/复氧组、薯蓣皂苷低、中、高剂量(10,20,40μmol/ml)干预组。MTT法检测心肌细胞存活率;TUNEL法检测心肌细胞凋亡百分率;Fluo-3负载激光扫描共聚焦显微镜观察细胞内[Ca2+]i的变化;硝酸还原酶法测定心肌细胞培养液中一氧化氮(NO)浓度。结果:与缺氧/复氧组比较,薯蓣皂苷各剂量干预组心肌细胞存活率明显升高,细胞凋亡率明显降低,心肌细胞[Ca2+]i荧光强度明显减弱,心肌细胞培养液中NO浓度降低。结论:薯蓣皂苷能有效抑制缺氧/复氧导致的乳鼠心肌细胞的凋亡,其可能与减轻细胞内钙超载,抑制受损心肌细胞NO升高,减轻过量NO的细胞毒性作用有关。

Objective：To study the effects of dioscin on cardiomyocytes apoptosis after anoxia/reoxygenation（A/ R） injury and the possible mechanism.Methods：Cultured myocardial cells of neonatal Wistar rats were randomly divided into five groups：control group;A/R group;dioscin L（10μmol/ml）,M（20μmol/ml）,H（40μmol/ml） group.In this experiment,the cellular livability was measured by MTT assay;the apoptosis percentage was measured by TUNEL;the intracellular calcium level was observed by laser scanning confocal microscopy marked Fluo-3/ AM;The concentrations of nitric oxide（NO） in cardiomyocytes cultured fluid were determined with nitrate reductase.Result：Compared with A/R group,The cellular livability of the treatment groups were significantly increased,the apoptosis percentage and the fluorescent intensity of intracellular calcium were significantly decreased,and the concentrations of NO was decreased.Conclusion：Dioscin has notable effects on cardiomyocytes apoptosis after anoxia-reoxygenation injury,which may be partly due to its alleviviating intracellular calcium overload,inhibiting the concentration of NO elevating in damaged cardiomyocytes and decreasing cellular toxic effects of high concentration of NO.