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兩種細胞建立肝細胞氧化損傷模型比較

發(fā)布時間:2018-02-22 05:54

  本文關鍵詞: 過氧化氫(HO) 氧化損傷 人肝癌細胞株(Hep G) 正常肝細胞株(Chang liver) 出處:《中國公共衛(wèi)生》2015年03期  論文類型:期刊論文


【摘要】:目的比較過氧化氫誘導人肝癌細胞株(Hep G2)與正常肝細胞株(Chang liver)建立的肝細胞氧化應激損傷模型。方法用不同濃度過氧化氫誘導Hep G2細胞和Chang liver細胞氧化應激損傷,采用噻唑藍法檢測細胞生長抑制率;分光光度法測定培養(yǎng)液中乳酸脫氫酶(LDH)、谷丙轉氨酶(ALT)、谷草轉氨酶(AST)活性,以及肝細胞中超氧化物歧化酶(SOD)活性、還原型谷胱甘肽(GSH)及丙二醛(MDA)含量。結果作用時間在0.5~4 h時,在75~600μmol/L濃度范圍內(nèi),過氧化氫可濃度和時間依賴性地抑制2種肝細胞增殖,促使細胞內(nèi)AST、ALT和LDH向培養(yǎng)液中釋放;細胞中SOD和GSH活性明顯降低,MDA含量明顯升高,表明2種肝細胞氧化損傷模型構建成功;選擇300μmol/L H2O2作用4 h為致Hep G2和Chang liver細胞氧化應激損傷的最佳條件,結果顯示,Hep G2和Chang liver細胞的生長抑制率分別為62%和76%,培養(yǎng)液中ALT、AST、LDH活性分別為(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,細胞中MDA含量分別為(7.8±0.9)和(8.6±1.1)nmol/mgprot。結論Hep G2與Chang liver細胞均可用于氧化應激細胞損傷模型的制備。
[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 鹵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 鹵0. 9 and 8. 6 鹵1. 1 nmol / mg prot, respectively.
【作者單位】: 延邊大學醫(yī)學院生物化學與分子生物學教研室;
【基金】:國家自然科學基金(81160539;81360651)
【分類號】:R575

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