丹参酮ⅡA下调氧化应激心肌细胞内PDCD5表达

TanshinoneⅡA down-regulates the expression of PDCD5 in cardiomyocytes with oxidative stress

目的:研究程序性死亡因子5(programed cell death5,PDCD5)在H2O2致心肌细胞氧化应激损伤中的表达及中药丹参酮ⅡA对其调节作用。方法:以H2O2干预心肌细胞,建立氧化应激心肌损伤模型;MTT法检测心肌细胞损伤程度;流式细胞术检测细胞凋亡率;AnnexinV-FITC染色法观察细胞凋亡形态;免疫荧光法检测PDCD5基因的表达。结果:与正常组相比,模型组的细胞活力明显下降,凋亡率显著增高,差异有统计学意义(P<0.01);丹参酮ⅡA组的细胞活力较模型组明显上升,凋亡率显著降低,其中中剂量组尤为明显,与模型组相比差异有统计学意义(P<0.01);PDCD5在正常心肌细胞内表达较低,而在模型组表达明显增高,在丹参酮ⅡA用药组其表达有所降低,中剂量组降低更明显,与模型组相比有显著性差异(P<0.05)。结论:H2O2可明显增加心肌细胞PDCD5的表达水平,丹参酮ⅡA可通过抑制该基因的过度表达起到减少细胞凋亡保护心肌细胞的作用。

Objective：To study the expression of PDCD5 in cardiomyocytes treated with H2O2 and the regulation of tanshinoneⅡA on PDCD5.Methods：Cardiomyocytes were treated with H2O2 to establish the oxidative stress model.The cadiomyocytes viability was detected by MTT.The ratio of apoptosis was checked out by flow cytometry.The morphous of apoptotic cells was presented by staining with AnnexinV-FITC.The expression of PDCD5 was measured by immunofluorescence.Results：The cell viability in model group was lower than that in control.The ratio of apoptosis in model group was obviously higher than that in control.There were significantly differences between the two groups（P〈0.01）.Pre-treating cardiomyocytes with tanshinoneⅡA for 24h could not only enhance the cell viability,but also decrease the ratio of apoptosis induced by H2O2,especially in medium dose group.It was significantly different from the model group（P〈0.01）.The expression level of PDCD5 in control group was very low,while it had an obvious increase in model.The fluorescence intensity in control was about 12.35±2.1,while that in model 64.28±4.9.TanshinoneⅡA decreased the expression of PDCD5 in cardiomyocytes treated with H2O2.The reduction in medium dose of tanshinoneⅡA group was the most obvious.There was significantly difference between the medium dose group and the model one（P〈0.05）.Conclusion：H2O2 obviously increases the expression of PDCD5 in cardiomyocytes.TanshinoneⅡA decreases the ratio of apoptosis and protects cardiomyocytes from the damage induced by H2O2 through down-regulation of the level of PDCD5.