两种细胞建立肝细胞氧化损伤模型比较

发布时间：2018-02-22 05:54

本文关键词： 过氧化氢(HO) 氧化损伤人肝癌细胞株(Hep G) 正常肝细胞株(Chang liver)　出处：《中国公共卫生》2015年03期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:Objective to compare the oxidative stress injury model of Hep G2 cells and Chang liver cells induced by hydrogen peroxide (Hep G 2) and normal liver cell line Chang liver.Methods the oxidative stress injury of Hep G2 cells and Chang liver cells was induced by hydrogen peroxide. The cell growth inhibition rate was measured by thiazolyl blue assay, and the activities of lactate dehydrogenase (LDH), alanine aminotransferase (alt), aspartate aminotransferase (AST), and superoxide dismutase (SOD) activity in hepatocytes were measured by spectrophotometry. The contents of reduced glutathione (GSH) and malondialdehyde (MDA) were found to be in the range of 75 ~ 600 渭 mol/L for 4 h. Results hydrogen peroxide could inhibit the proliferation of two kinds of hepatocytes in a dose-and time-dependent manner and promote the release of alt and LDH into the culture medium. The activity of SOD and GSH in the cells decreased significantly, which indicated that the two models of oxidative injury of hepatocytes were successfully constructed, and the best condition for oxidative stress injury of Hep G2 and Chang liver cells was to select 300 渭 mol/L H 2O 2 for 4 h. The results showed that the growth inhibition rates of Hep G2 and Chang liver cells were 62% and 76, respectively. The activities of Hep G 2 and Chang liver were 18.2 卤0.2nmol / mg prot. Conclusion Hep G2 and Chang liver cells could be used to prepare the model of oxidative stress cell injury. The results showed that the MDA content of Hep G2 and Chang liver cells were 62% and 76, respectively. Conclusion both Hep G2 and Chang liver cells can be used to prepare the model of oxidative stress cell injury. The results showed that the MDA content of Hep G 2 and Chang liver cells were 62% and 76, respectively. Conclusion both Hep G2 and Chang liver cells can be used to prepare the model of oxidative stress cell injury. The results showed that the MDA content of Hep G 2 and Chang liver cells were 7. 8 卤0. 9 and 8. 6 卤1. 1 nmol / mg prot, respectively.
【作者单位】：延边大学医学院生物化学与分子生物学教研室;
【基金】：国家自然科学基金(81160539;81360651)
【分类号】：R575

【相似文献】