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目的:探讨淫羊藿总黄酮(EF)与吡柔比星(THP)联用体外抗人骨肉瘤MG-63细胞的作用机制。方法:设立空白对照组和EF组、THP组及两药联合组4个实验组,倒置相差显微镜观察各组不同时间段细胞形态;作用48 h后,采用CCK-8法检测各组不同浓度的细胞抑制率,计算两药联用的联合指数(CI),分析其相互作用;采用Annexin V-FITC/PI双染法流式细胞术检测各组细胞凋亡情况;RT-PCR及Western-blotting检测Caspase-3、P21 mRNA及其蛋白的表达。结果:实验组细胞呈不同程度凋亡状态,单药组细胞增殖抑制率随药物浓度增加呈递增趋势,联合组细胞增殖抑制率及凋亡率均明显高于两单药组,联合组抑制率在10%~90%时CI值处于0.374~0.770之间,EF和THP均能上调Caspase-3、P21 mRNA及Caspase-3、p21蛋白的表达,二者联合上调趋势更明显。结论:EF和THP通过诱导细胞凋亡并明显抑制骨肉瘤MG-63细胞活性,二者联合相互作用呈现协同效应。
Objective: To investigate the mechanism of action of Epimedium sagittatum flavonoids (EF) and pirarubicin (THP) in vitro on human osteosarcoma MG-63 cells. Methods: The control group and the EF group, the THP group and the two drug combination group of four experimental groups, inverted phase contrast microscope observation of the different time periods of the cell morphology; after 48 h, CCK-8 method was used to test the different concentrations of each group Cell inhibition rates were calculated, the combination of the two drugs with the joint index (CI), analysis of their interaction; Annexin V-FITC / PI double staining flow cytometry was used to detect apoptosis in each group; RT-PCR and Western-blotting The expressions of Caspase-3, P21 mRNA and their proteins were detected. Results: The cells in the experimental group showed different degree of apoptosis. The inhibition rate of proliferation in the single drug group increased with the increase of the drug concentration. The proliferation inhibition rate and apoptosis rate in the combined group were significantly higher than those in the two single drug group. The inhibition rate CI values ranged from 0.374 to 0.770 at 10% -90%. Both EF and THP upregulated the expression of Caspase-3, P21 mRNA and Caspase-3, p21 protein. CONCLUSION: EF and THP show a synergistic effect by inducing apoptosis and significantly inhibiting MG-63 cell activity.