Abstract:
ObjectiveTo compare the utilization of HepG2 and Chang liver cell lines in hydrogen peroxide(H
2O
2)-induced hepatotoxicity cell models
in vitro.
MethodsMethylthiazolyldiphenyl-tetrazolium bromide(MTT)assay was used to evaluate the inhibitory effect of H
2O
2 on cell proliferation.Lactate dehydrogenase(LDH),alanine amino-transferase(ALT),and aspartate aminotransferase(AST)in culture solution and malondialdelyde(MDA),superoxide dismutase(SOD),and reduced glutathione(GSH)in the cells were detected with spectrophotometric method.
ResultsH
2O
2 at the concentrations of 75-600 μmol/L and with the treatment time of 0.5 to 4 hours inhibited the proliferation of the two cell lines and resulted in the leakage of cellular LDH,ALT and AST into culture solution in a concentration- and time-dependent manner;furthermore,H
2O
2 increased MDA formation and reduced GSH level in the cells of the two cell lines.The results suggested that H
2O
2 could induce cellular oxidative injury in both HepG2 and Chang liver cell line.With the optimal H
2O
2 concentration of 300 μmol/L and treatment time of 4 hours,the proliferation inhibitory ratio,the activities of ALT,AST and LDH in the culture solution,and the concentration of MDA in the cells were 62%,18.2±0.2 U/L,34.2±4.6 U/L and 544.2±26.8 U/L,and 8.6±1.1 nmol/mg prot for the model utilizing HepG2 cell line,and those indicators were 76%,19.1±0.1 U/L,30.3±2.5 U/L and 536.8±22.3U/L,and 7.8±0.9 nmol/mg prot for the model utilizing Chang liver cell line.
ConclusionThe two cell lines could be utilized in the construction of H
2O
2-induced cellular oxidative damage.