Establishment of cell injury model by H2O2-induced oxidative stress in human melanocytes

doi:10.11786/sypfbxzz.1674-1293.20140603

Abstract：ObjectiveTo establish an oxidative stress injury model of PIG1 cells with different concentrations of H2O2 and to further study the mechanism of oxidative stress in vitiligo.Methods After 24 hour treatment with different concentrations of H2O2, the morphological changes of PIG1 cells were observed by light microscopy. The intracellular ROS and apoptosis ratio of PIG1 cells were determined by lfow cytometry method. The cell viability was determined by CCK-8 assay. The malondialdehyde (MDA) content in PIG1 was determined by measuring thiobarbituric acid reactive substances to monitor lipid peroxidation.Results With the increasing of the concentration of H2O2, melanocytes turned smaller; the number of dendrites decreased and the length of them became shorter; cell death (apoptosis and necrosis) increased gradually. The apoptosis ratio of PIG1 cells induced by 0.50, 0.75, 1.00, 1.25, 1.50 and 2.00 mmol/L H2O2 were 6.8%, 11.5%, 15.6%, 22.7%, 38.7% and 57.5% respectively. Compared to the 0 mmol/L group, the viability of PIG1 cells was signiifcantly decreased when cells were treated by 1.0 mmol/L H2O2(P<0. 05). Compared with the control group, the intracellular ROS and the MDA content was signiifcantly elevated when cells were treated by 1.0 mmol/L H2O2(P<0.05). Conclusion The optimal concentration of H2O2 which could induce oxidative stress in PIG1 cells was determined and the cell injury model induced by H2O2 was successfully established.