葛根素对过氧化氢诱导平滑肌细胞凋亡的保护作用

Protective effect of puerarin on vascular smooth muscle cell apoptosis induced by H_2O_2

目的探讨葛根素对过氧化氢(双氧水)H2O2诱导大鼠胸主动脉平滑肌细胞凋亡保护作用的机制。方法在过氧化氢(双氧水)H2O2诱导大鼠胸主动脉平滑肌细胞凋亡模型的基础上,以噻唑兰(MTT)法测定细胞存活率;用流式细胞术和Hoechst33258荧光染色法观察细胞受损和凋亡;用DNA琼脂糖凝胶电泳法观察细胞凋亡过程中DNA断裂程度;用免疫细胞组织化学染色法检测细胞内Caspase-3的表达。结果过氧化氢(双氧水)H2O2诱导无血清培养的平滑肌细胞凋亡,葛根素可显著降低平滑肌细胞的凋亡,减少凋亡细胞DNA断裂,平滑肌细胞坏死减少,对Caspase-3的表达有明显抑制作用。结论葛根素对过氧化氢(双氧水)H2O2诱导血管平滑肌细胞凋亡有保护作用,其机制可能与抑制Caspase-3有关。

[Objective] To investigate the mechanism and the protective effects of puerarin on the smooth muscle cells apoptosis induced by hydrogen peroxide （H2O2）. [Methods] Following the estabilishment of a model of the H2O2-indueed vascular smooth muscle cells apotosis, the apoptosis of the cells were measured by floweytometry and Hoechst 33258 staining; the DNA fragmentation was analyzed by agarose gel electrophorsis; the viability percentage of cells was measured using MTl assay. The expression of Caspase-3 in endothelial cells was detected by immuno- eytoehemieal method. [Results] The apoptosis of VSMC occurred in serum free medium （18 h） containing H2O2. When the VSMC had been treated with puerarin, the percentage of VSMC apoptosis was markedly decreased, and the degraded fragments of DNA obviously decreased. Puerarin （10^-3, 10^-4, 10^-5molF/L） could partly protect cells from neerosisi induced by H2O2 and reduce the expression of Caspase-3 significantly. [Conclusion] Puerarin can protect apoptotic vascular smooth muscle cells induced by H2O2 markedly and the effect is performed partly by decreasing Caspase-3 expression.