二苯乙烯苷对氧化诱导内皮细胞凋亡及Caspase-3和PARP表达的影响

Effects of TSG on Apoptosis of HUVECs and The Expression of Caspase-3 and PARP Induced by H/_2O_2

为研究二苯乙烯苷对过氧化氢(H2O2)诱导人脐静脉内皮细胞凋亡的影响并初步探讨二苯乙烯苷抗凋亡的可能机制,采用不同浓度H2O2和二苯乙烯苷处理内皮细胞24 h,以MTT法检测细胞生长活力、Hoechst33258染色观察细胞形态、流式细胞仪检测凋亡率等方法筛选造模内皮细胞凋亡的H2O2浓度和二苯乙烯苷最佳的抗内皮细胞凋亡作用浓度.RT-PCR、Western-blot分别检测Caspase-3、PARP mRNA及蛋白质的表达.结果发现,与空白对照组相比,300μmol/L H2O2作用后,内皮细胞增殖明显受到抑制,Hoechst33258染色可见大量凋亡细胞,细胞凋亡率显著增加,流式细胞仪检测出明显的凋亡峰,Caspase-3和多聚腺苷酸二磷酸核糖基聚合酶(PARP)的表达量显著增加.经二苯乙烯苷处理后,随着二苯乙烯苷浓度增加,细胞的增殖率随之增加,凋亡细胞数减少,凋亡率逐渐降低,与H2O2组比较,10μmol/L的二苯乙烯苷能够显著提高细胞增殖率,降低细胞凋亡率,并显著减少Caspase-3和PARP表达.以上结果表明,二苯乙烯苷能够抑制由H2O2诱导的人脐静脉内皮细胞凋亡,增加细胞生长活性,降低细胞凋亡率,其作用机制可能与下调Caspase-3和多聚腺苷酸二磷酸核糖基聚合酶(PARP)的表达有关.

To explore the effects of tetrahydroxystilbene-2-O-β-D-glucoside（TSG） on the apoptosis of HUVEC cells and its mechanisms induced by H2O2.The models of apoptosis induced by H2O2 in HUVEC and the concentrations of TSG on apoptosis model were established via MTT,Hoechst33258 staining,and Flow Cytometry.The expression of Caspase-3 and PARP was detected by RT-PCR and Western blot.Compared with control group,the viability of cells is decreased and the apoptosis ratios is increased with increased concentration of H2O2（P 0.01） respectively.And the group of 300 μmol/L H2O2 inhibited the cell proliferation,increased the number of apoptotic cells significantly.The viability of cells is increased and the apoptosis of HUVECs is decreased after pretreated with different concentration of TSG.According to the MTT and Flow Cytometry,the optimal concentration for H2O2 to establish apoptosis model and for TSG to protect HUVECs induced by H2O2 were 300 μmol/L and 10 μmol/L respectively.Compared with the control group,the group of 300 μmol/L H2O2 inhibited the cell proliferation,increased the number of apoptotic cells and the expression of Caspase-3 significantly.Compared with H2O2 group,10 μmol/L of TSG improved the rate of cell proliferation,inhibited cell apoptosis,and decreased the expressions of the Caspase-3 and PARP significantly（P 0.01）.These results indicate that TSG can inhibit H2O2-induced apoptosis of human umbilical vein endothelial cells,and its mechanism may relate to the down-regulation of Caspase-3 and PARP expression.