两种细胞建立肝细胞氧化损伤模型比较

目的比较过氧化氢诱导人肝癌细胞株(HepG2)与正常肝细胞株(Chang liver)建立的肝细胞氧化应激损伤模型。方法用不同浓度过氧化氢诱导HepG2细胞和Chang liver细胞氧化应激损伤, 采用噻唑蓝法检测细胞生长抑制率;分光光度法测定培养液中乳酸脱氢酶(LDH)、谷丙转氨酶(ALT)、谷草转氨酶(AST)活性, 以及肝细胞中超氧化物歧化酶(SOD)活性、还原型谷胱甘肽(GSH)及丙二醛(MDA)含量。结果作用时间在0.5~4 h 时, 在75~600 μmol/L浓度范围内, 过氧化氢可浓度和时间依赖性地抑制2种肝细胞增殖, 促使细胞内AST、ALT和LDH向培养液中释放;细胞中SOD和GSH活性明显降低, MDA含量明显升高, 表明2种肝细胞氧化损伤模型构建成功;选择300 μmol/L H₂O₂作用4 h为致HepG2和Chang liver细胞氧化应激损伤的最佳条件, 结果显示, HepG2和Chang liver细胞的生长抑制率分别为62%和76%, 培养液中ALT、AST、LDH活性分别为(18.2±0.2)、(34.2±4.6)、(544.2±26.8)和(19.1±0.1)、(30.3±2.5)、(536.8±22.3)U/L, 细胞中MDA含量分别为(7.8±0.9)和(8.6±1.1)nmol/mgprot。结论HepG2与Chang liver细胞均可用于氧化应激细胞损伤模型的制备。

Comparative study on utilization of two cell lines in liver cell oxidative damage model

ObjectiveTo compare the utilization of HepG2 and Chang liver cell lines in hydrogen peroxide(H₂O₂)-induced hepatotoxicity cell models in vitro.MethodsMethylthiazolyldiphenyl-tetrazolium bromide(MTT)assay was used to evaluate the inhibitory effect of H₂O₂ 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.ResultsH₂O₂ 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,H₂O₂ increased MDA formation and reduced GSH level in the cells of the two cell lines.The results suggested that H₂O₂ could induce cellular oxidative injury in both HepG2 and Chang liver cell line.With the optimal H₂O₂ 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 H₂O₂-induced cellular oxidative damage.