马鞭草总黄酮诱导肝癌HepG-2细胞凋亡及可能机制

Apoptosis of HepG-2 cells induced by total flavonoids of Verbena officinalis L. and possible mechanism

目的探讨马鞭草总黄酮对肝癌HepG-2细胞凋亡的作用及可能机制。方法采用浓度为50、100和200 mg/L的马鞭草总黄酮处理体外培养的肝癌HepG-2细胞,流式细胞仪和琼脂糖凝胶电泳检测细胞凋亡;流式细胞仪检测活性氧簇(ROS)和膜电位变化;Western印迹法分析Caspase-3、Caspase-8、Caspase-9和P53蛋白表达水平。结果与未给药对照组比较,马鞭草总黄酮50、100和200 mg/L给药组均能促进肝癌HepG-2细胞凋亡(P<0.01),增加ROS水平(P<0.01),降低线粒体膜电位(P<0.05,P<0.01);马鞭草总黄酮50、100和200 mg/L可增加Caspase-3、Caspase-9和P53蛋白表达水平(P<0.05,P<0.01)。结论马鞭草总黄酮可体外诱导肝癌HepG-2细胞凋亡,其诱导凋亡的机制可能与其增加ROS水平、降低线粒体膜电位,并同时上调Caspase-3、Caspase-9、P53蛋白水平有关。

Objective To explore the apoptosis and mechanisms of HepG-2 cells induced by total flavonoids of Verbena officinalis L.（TFV）. Methods HepG-2 cells were cultured with different concentrations of TFV. The apoptosis of HepG-2 cells was evaluated by flow cytometry and DNA ladder. Reactive oxygen species（ROS）and membrane potential were measured by flow cytometry. Expression of Caspase-3,Caspase-8,Caspase-9,and P53 was analyzed by Western blotting. Results TFV 50,100 and 200 mg/L increased the apoptosis rate（P〈0.01）,increased ROS levels of HepG-2 cells（P〈0.01）,and decreased the mitochondria membrane potential（P〈0.05,P〈0.01）. TFV（50,100 and 200 mg/L）increased the proteins′ expression of Caspase-3,Caspase-9 and P53（P〈0.05,P〈0.01）. Conclusion TFV may induce the apoptosis of HepG-2 cells via increasing ROS levels,decreasing mitochondria membrane potential,and up-regulateing the expression of Caspase-3,Caspase-9 and P53 protein in HepG-2 cells.