丹参酮ⅡA对Adriam诱导的人肾小球系膜细胞凋亡的保护作用

Protective effects of tanshinone ⅡA on adriamycin-induced apoptosis of human mesangial cells

目的探究丹参酮ⅡA (TSⅡA)对阿霉素(Adriam)诱导的肾小球系膜细胞HMCL凋亡的作用。方法用不同浓度的TSⅡA处理HMCL细胞,确定安全用药范围;将细胞随机分为HMCL组、Adriamycin组、Adriam+TSⅡA(5μM)组、Adriam+TSⅡA(10μM)组和Adriam+TSⅡA(20μM)组。除HMCL组外,其余组均用Adriamycin处理细胞,Adriam+TSⅡA(5,10,20μM)组同时加入5、10、20μM的TSⅡA处理细胞,用CCK8法检测细胞增殖,流式和Hoechst染色检测细胞凋亡,Western blot检测Caspase-3和Caspase-9的表达。结果与HMCL组比较,Adriamycin组细胞增殖速度明显降低;与Adriamycin组比较,Adriam+TSⅡA (5,10,20μM)组细胞增殖速度明显升高;同时,Adriamycin组细胞凋亡率与HMCL组比较明显升高,Adriam+TSⅡA (5,10,20μM)组细胞凋亡率明显低于Adriamycin组;此外,Adriam能显著促进HMCL细胞Caspase-3和Caspase-9表达,TSⅡA能显著减弱Adriam促进Caspase-3和Caspase-9表达的作用。结论 TSⅡA能通过抑制凋亡相关蛋白的表达抑制Adriam诱导的肾小管系膜细胞凋亡。

Objective To investigate the effects of tanshinone ⅡA （TS ⅡA） on adriamycin （Adriam）-induced apoptosis of human mesangial cells HMCL. Methods Cells were treated with different concentrations of TS ⅡA to determine the safe range on HMCL.Cells were divided into HMCL group,Adriamycin group,Adriam＋ TSⅡA （5 μM） group,Adriam＋ TSⅡA （10 μM） group and Adriam＋ TSⅡA （ 20 μM） group and treated with Adriamycin except HMCL group.Adriam＋ TSⅡA （5,10,20 μM） group also were treated with TSⅡA （5,10,20 μM） respectively.Cell proliferation was measured by CCK8 assay,flow cytometry and Hoechst staining were performed for cell apoptosis.The expression levels of Caspase-3 and Caspase-9 were determined by Western blot. Results Compared with HMCL group, cell growth was inhibited in Adriamycin group;compared with Adriamycin group,cell growth was induced in Adriam＋ TSⅡA （5,10,20 μM） group.Meanwhile,the apoptosis rate in Adriamycin group was increased compared with HMCL group,but decreased in Adriam＋ TSⅡA （5,10,20 μM） group compared with Adriamycin group.In addition,Adriam induced expressions of Caspase-3 and Caspase-9. TSⅡA attenuated the effects of Adriam on Caspase-3 and Caspase-9. Conclusion TSⅡA inhibits Adriam-induced apoptosis of mesangial cells via inhibiting expressions of apoptosis-related proteins.