This study investigated the effect of arctigenin(ARG) on the induction of apoptosis and the putative pathways of its action in HL-60 and K562 cells.MTT assay was used to detect the cytotoxic effect of ARG in HL-60 and...This study investigated the effect of arctigenin(ARG) on the induction of apoptosis and the putative pathways of its action in HL-60 and K562 cells.MTT assay was used to detect the cytotoxic effect of ARG in HL-60 and K562 cells.The apoptosis was observed by Hoechst 33258 fluorescence staining and DNA agarose gel electrophoresis.Caspase-3 enzyme activity was measured by caspase-3 enzyme activity detection kit.The expression of related protein was analyzed by Western blotting and the vascular endothelial growth factor(VEGF) level was determined by enzyme-linked immunosorbent assay(ELISA).ARG-treated HL-60 cells and K562 cells exhibited growth inhibition and displayed several features of apoptosis,including DNA fragmentation and DNA laddering by agarose gel electrophoresis.It was observed that poly-(ADP-ribose) polymeras(PARP) were cleaved to smaller molecules and ARG induced upregulation of bax and downregulation of bcl-2 protein expression.However,it had no effect on VEGF levels.Taken together,this study demonstrated that ARG is a potent inducer of apoptosis and this was accompanied by caspase-3 activation and upregulation of bax/bcl-2,which offers a potential mechanism for the apoptosis-inducing activity of ARG.展开更多
文摘建立了分离鉴定牛蒡叶中微量木脂体牛蒡子甙(arctiin)和牛蒡子甙元(arctigenin)的方法。牛蒡叶粗提物经聚酰胺柱提取,浓缩其甲醇洗脱液并于低温下静置析出白色沉淀物。沉淀物用甲醇溶解后经反相高效液相色谱(RP HPLC)分离,纯化得到两个主要组分。经紫外光谱(UV)、红外光谱(FTIR)及电喷雾质谱(ESI MS)检测,并与牛蒡子甙和牛蒡子甙元对照品的UV,LC ESI MS,HPLC及FTIR图谱比较,鉴定两个主要成分为牛蒡子甙和牛蒡子甙元。
文摘This study investigated the effect of arctigenin(ARG) on the induction of apoptosis and the putative pathways of its action in HL-60 and K562 cells.MTT assay was used to detect the cytotoxic effect of ARG in HL-60 and K562 cells.The apoptosis was observed by Hoechst 33258 fluorescence staining and DNA agarose gel electrophoresis.Caspase-3 enzyme activity was measured by caspase-3 enzyme activity detection kit.The expression of related protein was analyzed by Western blotting and the vascular endothelial growth factor(VEGF) level was determined by enzyme-linked immunosorbent assay(ELISA).ARG-treated HL-60 cells and K562 cells exhibited growth inhibition and displayed several features of apoptosis,including DNA fragmentation and DNA laddering by agarose gel electrophoresis.It was observed that poly-(ADP-ribose) polymeras(PARP) were cleaved to smaller molecules and ARG induced upregulation of bax and downregulation of bcl-2 protein expression.However,it had no effect on VEGF levels.Taken together,this study demonstrated that ARG is a potent inducer of apoptosis and this was accompanied by caspase-3 activation and upregulation of bax/bcl-2,which offers a potential mechanism for the apoptosis-inducing activity of ARG.
