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目的比较过氧化氢诱导人肝癌细胞株(Hep G2)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Hep G2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75~600μmol/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300μmol/L H2O2作用4 h为致Hep G2和Chang liver细胞氧化应激损伤的最佳条件,结果显示,Hep G2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Hep G2与Chang liver细胞均可用于氧化应激细胞损伤模型的制备。
Objective To compare the hepatocellular oxidative stress injury model induced by hydrogen peroxide (Hep G2) and normal liver cell (Chang liver). Methods Oxidative stress injury was induced in Hep G2 cells and Chang liver cells by different concentrations of hydrogen peroxide. Cell growth inhibition rate was measured by thiazolyl blue assay. The levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT) ), Aspartate aminotransferase (AST) activity, superoxide dismutase (SOD) activity, reduced glutathione (GSH) and malondialdehyde (MDA) Results Hydrogen peroxide could inhibit the proliferation of two kinds of hepatocytes in a concentration-dependent manner in a concentration range of 75-600 μmol / L for 0.5-4 h, and promote the release of intracellular AST, ALT and LDH ; The activities of SOD and GSH in the cells were significantly decreased and the content of MDA was significantly increased, indicating that the oxidative damage model of two kinds of hepatocytes was successfully established. Selecting 300μmol / L H2O2 for 4 h was the best oxidative stress injury to Hep G2 and Chang liver cells The results showed that the growth inhibition rates of Hep G2 and Chang liver cells were 62% and 76%, respectively. The activities of ALT, AST and LDH in culture medium were (18.2 ± 0.2), (34.2 ± 4.6) and (544.2 ± 26.8) ) And (19.1 ± 0.1), (30.3 ± 2.5) and (536.8 ± 22.3) U / L respectively. The MDA contents in the cells were (7.8 ± 0.9) and (8.6 ± 1.1) nmol / mgprot, respectively. Conclusion Both Hep G2 and Chang liver cells can be used in the model of oxidative stress cell injury.