黄芪甲苷对大鼠肝星状细胞氧化损伤的作用研究

Effect of Astragalus Saponin on Oxidative Damage in Rats with Hepatic Stellate Cells

目的探讨黄芪甲苷对H_2O_2诱导大鼠肝星状细胞氧化损伤的作用及相关机制。方法将大鼠肝星状细胞分为对照组、氧化损伤组(100μmol/L的H_2O_2)、黄芪甲苷(100μmol/L、200μmol/L)干预组。应用甲基噻唑基四唑(MTT)比色法检测肝星状细胞增殖率,流式细胞术测定肝星状细胞内活性氧(ROS)表达,Hoechst-PI染色观察细胞凋亡情况,比色法测定细胞内Caspase-3及Caspase-9的表达水平。结果黄芪甲苷干预组肝星状细胞活性高于氧化损伤组,肝星状细胞内ROS水平、凋亡细胞比例和细胞内Caspase-3及Caspase-9表达水平低于氧化损伤组(P<0.05,P<0.01)。结论黄芪甲苷预处理可减轻大鼠肝星状细胞氧化损伤,其作用机制可能与抑制Caspase-3及Caspase-9的表达,从而抑制细胞凋亡有关。

Objective To investigate effect and mechanism of Astragalus Saponin on oxidative damage in rats with hepatic stellate cells induced by H 2O 2. Methods Rat hepatic stellate cells were divided into control group, oxidative damage group （100 μmol/L H 2O 2） and Astragalus Saponin （100 μmol/L, 200 μmol/L） intervention group. Methyl thiazolyl tetrazolium （MTT） colorimetry was used to detect proliferation rate of hepatic stellate cells, and flow cytometry technology was used to detect expression of reactive oxygen species （ROS） in hepatic stellate cells, and Hoechst-PI staining was used to observe apoptosis, and colorimetric method was used to determine expressions of Caspase-3 and Caspase-9 in cells. Results Activity value of hepatic stellate cells in Astragalus Saponin intervention group was significantly higher, while values of ROS level, apoptosis percentage and Caspase-3 and Caspase-9 expressions in hepatic stellate cells were significantly lower than those in oxidative damage group （ P 〈0.05, P 〈0.01）. Conclusion Pretreatment of Astragalus Saponin may decrease oxidative damage of stellate cells in rats, and its mechanism may be relate to inhibition of expressions of Caspase-3 and Caspase-9 to inhibit cells apoptosis.