过氧化氢诱导BRL细胞凋亡损伤模型的建立

Establishment of BRL cell apoptosis models by hydrogen peroxide

动物冷应激过程中肝脏因发生氧化应激而受损。本试验采用过氧化氢(H2O2)诱导buffalo大鼠肝细胞(BRL)凋亡,建立体外氧化应激的凋亡损伤模型,为深入研究冷应激对肝脏损伤的机理奠定基础。试验采用不同浓度H2O2刺激BRL细胞2h,运用WST-1法检测细胞生长活力、Annexin-V/FITC-PI双染技术检测细胞凋亡率及Western blot方法检测Caspase-3的表达量来筛选BRL细胞凋亡的H2O2作用浓度。结果显示,与空白对照组相比,150μmol/L H2O2作用后,细胞增殖率明显下降,Caspase-3的蛋白表达量增加,流式细胞仪检测的凋亡率升高,同时细胞坏死较其他H2O2处理组低。结果表明,150μmol/L H2O2作用BRL细胞2h可模拟氧化应激造成的凋亡损伤。

The animal liver is damaged by the occurrence of oxidative stress during cold stress.In the study,hydrogen peroxide(H2O2)was used to treat buffalo rat liver cells(BRL)to establish the apoptosis model indsced by in vitro oxidative stress.Different concentrations of H2O2 were used to stimulate BRL cells for 2h,then assayed the viability and apoptosis of BRL cells by WST-1,Annexin-V/FITC-PI double staining,and the protein expression of Caspase-3were analyzed using Western blot in order was to chose the optimal concentration of H2O2.Compared with the control group,the cell proliferation rate was significantly decreased,an increase in protein expression of Caspase-3and apopotosis was observed after treatment by 150μmol/L H2O2,but cell necrosis was lower than that of other H2O2 treatment.This study makes it possible to set up a cell model of apoptosis caused by H2O2 in vitro.