Establishment of Oxidative Damage Model of Bovine Mammary Epithelial Cells Induced by Hydrogen Peroxide

This study was conducted to establish the oxidative damage model of bovine mammary epithelial cells(BMEC) using hydrogen peroxide(H2O2) as stress source and using survival rate and antioxidant parameters as judgment indices.Experiment 1 was conducted as a single factor randomized arrangement,and the3 rd passage BMEC were divided into 42 groups with 10 replicates in each group.Cells were incubated in different concentrations of H2O2(0,100,200,400,600,800 and 1 000 μmol/L,respectively) for 2,4,6,8,12 and 24 h,respectively.The cell count was determined,and the survival rate was calculated.On the basis of experiment 1(optimal action time was 6 h),experiment 2 was also conducted as a single factor randomized arrangement,and the 3th passage BMEC were divided into 7 groups with 6 replicates in each group.Culture mediums containing different concentrations of H2O2(0,100,200,400,600,800 and 1 000 μmol/L,respectively) were used to act cell for 6 h.The cells and culture mediums were collected for the analysis of antioxidant parameters to select the appropriate concentration of H2O2 ensuring oxidative damage to cells.The results showed as follows: experiment 1,the BMEC survival rate reduced to 79.79% when the cells were induced by 600 μmol/L H2O2 for 6 h.Experiment 2,cell induced by 600 to 1 000 μmol/L H2O2 significantly decreased the activities of glutathione peroxidase,superoxide dismutase and catalase(P 0.05),and significantly increased malondialdehyde content compared with the other groups(P 0.05),but the differences among 600,800 and 1 000 μmol/L groups were not significant(P 0.10).The condition that 600 μmol/L H2O2 reacted for 6 h is optimal for establishment of oxidative damage model in BMEC.