Nrf2/ARE信号通路在槲皮素抑制异烟肼诱导的肝细胞线粒体氧化损伤中的作用

doi:10.3969/j.issn.1004-616x.2021.03.009

摘要/Abstract

摘要： 目的：探讨核因子E2相关因子2/抗氧化反应元件（Nrf2/ARE）信号通路在槲皮素抑制异烟肼（INH）诱导的L-02细胞线粒体氧化损伤中的作用。方法：将L-02细胞随机分为阴性对照组、INH组（10 mmol/L INH）、单独槲皮素组（50 μmol/L槲皮素）、槲皮素保护组（10 mmol/L INH+50 μmol/L槲皮素），各组细胞处理24 h后，采用四甲基偶氮唑蓝（MTT）法测定细胞存活率；荧光探针DCFH-DA检测细胞线粒体活性氧（ROS）水平；比色法测定细胞内丙二醛（MDA）含量、谷胱甘肽（GSH）含量和超氧化物歧化酶（SOD）活性，评价细胞线粒体氧化损伤状态。Western blot检测Nrf2、血红素氧合酶1（HO-1）蛋白表达水平，评价细胞内Nrf2/ARE信号通路的变化。结果：与阴性对照组比较，INH组细胞存活率显著降低（P < 0.01），线粒体ROS水平和MDA含量显著升高（P < 0.01），GSH含量及SOD活性明显降低（P < 0.01）；与INH组相比较，槲皮素保护组细胞存活率明显增加（P < 0.01），线粒体ROS水平和MDA含量显著减少（P < 0.01），GSH含量及SOD活性明显升高（P < 0.05或P < 0.01）。INH组细胞质中HO-1蛋白及细胞核中Nrf2蛋白表达较阴性对照组明显增加（P < 0.01）；槲皮素保护组细胞质中HO-1蛋白及细胞核中Nrf2蛋白表达明显高于INH组（P < 0.01）。结论：槲皮素能够抑制INH诱导的肝细胞线粒体氧化损伤，这可能与槲皮素对Nrf2/ARE信号通路的活性调节相关。

关键词: 槲皮素, 异烟肼, Nrf2/ARE信号通路, 线粒体, 氧化损伤

Abstract: OBJECTIVE： To investigate the role of the Nrf2/ARE signaling pathway on inhibitory effect of quercetin on mitochondrial oxidative damage which was induced by isoniazid (INH) in L- 02 hepatocytes. METHODS： L- 02 cells in cultures were randomly divided into several groups： negative control， INH (10 mmol/L)，quercetin alone (50 μmol/L) and combined (10 mmol/L INH+50 μmol/L quercetin). After cells were treated for 24 hours，cell viability was measured using the MTT method； levels of the mitochondrial reactive oxygen species (ROS) was detected by fluorescence probe DCFH-DA； contents of malondialdehyde (MDA)， glutathione (GSH) and the activity of superoxide dismutase (SOD) were measured using colorimetry to evaluate the oxidative damage status of mitochondria. In addition， the protein expressions of Nrf2 and HO-1 were detected using western blot analysis to assess changes in the Nrf2/ARE signaling pathway. RESULTS： Compared with the negative control group，cells from the INH group showed the followings：cell vitality was significantly decreased (P < 0.01)， mitochondrial ROS level and MDA content were significantly increased (P< 0.01)，GSH content and SOD activity were remarkable reduced (P < 0.01). Compared with the INH group，the other treated cells showed the followings： cell vitality was markedly increased (P < 0.01)， the level of mitochondrial ROS and the content of MDA were remarkable declined (P < 0.01)，the content of GSH and the activity of SOD were significantly elevated of the combined treatment group (P < 0.05 or P < 0.01). In addition， expressions of HO-1 protein in cytoplasm and Nrf2 protein in nucleus of the INH group were significantly higher than those in the negative control group (P < 0.01). Expressions of HO- 1 and Nrf2 proteins of the combined treatment group were markedly elevated compared with those of the INH group (P < 0.01). CONCLUSION： Quercetin inhibited mitochondrial oxidative damage which was induced by INH， and the mechanism might be mediated by quercetin-regulation of the Nrf2/ARE signaling pathway.

Key words: quercetin, isoniazid, Nrf2/ARE signaling pathway, mitochondrial, oxidative damage

中图分类号:

R965

陈廷玉, 陈大印, 谢金鹿, 高喜仁, 卢春凤. Nrf2/ARE信号通路在槲皮素抑制异烟肼诱导的肝细胞线粒体氧化损伤中的作用[J]. 癌变·畸变·突变, 2021, 33(3): 208-212,217.

CHEN Tingyu, CHEN Dayin, XIE Jinlu, GAO Xiren, LU Chunfeng. Role of the Nrf2/ARE signaling pathway on quercetin-inhibition of INH-induced mitochondrial oxidative damage in hepatocytes[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2021, 33(3): 208-212,217.

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