二苯乙烯苷对过氧化氢诱导内皮细胞凋亡的影响

Effect of 2,3,5,4'-Tetrahydroxystilbene-2-O-β-D-Glucoside on Hydrogen Peroxide-Induced Apoptosis of Human Umbilical Vein Endothelial Cell

目的探讨二苯乙烯苷对过氧化氢诱导人脐静脉内皮细胞凋亡的影响及其可能机制。方法体外培养人脐静脉内皮细胞,以过氧化氢作用内皮细胞建立细胞凋亡模型,再加入不同浓度的二苯乙烯苷作用24 h。采用MTT法检测细胞生长活力,Hoechst33258染色观察细胞形态,流式细胞仪检测细胞凋亡率,RT-PCR、免疫印迹法检测Caspase-3的表达。结果与空白对照组相比,300μmol/L过氧化氢作用后细胞增殖明显受到抑制,Ho-echst33258染色可见大量凋亡细胞,流式细胞仪检测出明显的凋亡峰,Caspase-3的表达量显著增加;加入二苯乙烯苷作用后,与过氧化氢组比较,10μmol/L二苯乙烯苷能够显著提高细胞增殖率,抑制细胞凋亡,并且使Caspase-3的表达减少(P<0.05)。结论二苯乙烯苷能够抑制过氧化氢诱导的人脐静脉内皮细胞凋亡,其作用机制与抑制Caspase-3表达有关。

Aim To investigate the role and mechanism of TSG on the apoptosis of human umbilical vein endothelial cells（HUVEC） induced by H2O2. Methods HUVEC were treated with TSG（0.1,1,10 and 100 μmol/L） for 24 hours then exposed to H2O2（100,200,300,400 and 500 μmol/L） for 24 hours,the optimal concentration of H2O2 and TSG were dedected by MTT and Flow Cytometry. Morphology of apoptosis and the protective effect of TSG on HUVEC induced by H2O2 were observed by Hoechst33258 staining.The mRNA and protein expression of Caspase-3 were detected by RT-PCR and Western blot analysis respectively. Results According to the MTT and Flow Cytometry,the optimal concentration for H2O2 to establish apoptosis model and for TSG to protect HUVEC 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 expression of the caspase-3 significantly（P0.05）. Conclusion TSG could inhibit H2O2-induced apoptosis of HUVEC,and its mechanism was associated with the inhibition of Caspase-3 expression.