管花肉苁蓉苯乙醇苷提取物对血管内皮细胞EA.hy926增殖的抑制和凋亡的诱导作用

Proliferation inhibition and apoptosis induction of phenylethanoid glycoside extracted from Cistanche tubulosa on EA. hy926 cells

目的研究管花肉苁蓉苯乙醇苷提取物对血管内皮细胞EA.hy926增殖的抑制和凋亡的诱导作用。方法采用CCK-8法测定细胞增殖的抑制率,用Hoechst33342染色法观察细胞的凋亡形态,采用Annexin V-FITC/PI双染法结合流式细胞仪测定细胞的凋亡率,用Western blot法检测凋亡相关蛋白Bcl-2、Bax的表达情况。结果与对照组相比,管花肉苁蓉苯乙醇苷提取物能够抑制血管内皮细胞的增殖且呈剂量依赖性,其作用24 h时的IC50=30.61μg·m L-1;能够增加血管内皮细胞凋亡率,药物组的细胞呈现核浓染和固缩现象,能够降低抗凋亡蛋白Bcl-2的表达、增加促凋亡蛋白Bax的表达、Bcl-2/Bax值显著降低。结论肉苁蓉苯乙醇苷提取物能够通过诱导血管内皮细胞的凋亡来抑制其增殖,其机制与调控凋亡相关蛋白Bcl-2、Bax的表达有关。

OBJECTIVE To investigate the effects of phenylethanoid glycoside from Cistanche tubulosa on proliferation and apoptosis of human umbilical vein endothelial cell line EA. hy926. METHODS EA. hy926 ceils were treated with different concentrations of phenylethanoid glycoside from Cistarwhe tubulosa. Inhibitory effects were measured by CCK - 8 assay. The morphological changes of apoptotic cells were determined using Hoechst33342 staining. The percentage of apoptotic ceils were detected by Annexin V - FITC/PI staining and flow cytometry. Furthermore, the expression of apoptosis - related proteins was determined by Western blot assay. RESULTS Phenylethanoid glycoside inhibited the proliferation of EA. hy926 cells in a concentration - dependent manner with an IC50 value of 30.61 μg·m L^-1. CellLs incubated with phenylethanoid glycoside showed the typical staining of apoptotic cells. After treatment with phenylethanoid glycoside,the percentage of apoptotic ceils increased significantly. Western blot showed that phenylethanoid glyco- side could decrease the ratio of Bcl - 2/Bax. CONCLUSION These results suggest that the inhibitory effects of phenylethanoid glyco- side on proliferation of EA. hy926 cells is the results of apoptosis. Phenylethanoid glycoside may be a potential apoptosis - inducing agent for endothelial cells.