摘要:
为探讨ROS介导的氧化应激在异烟肼(INH)诱导L-02细胞毒性中的作用及槲皮素的干预作用,建立体外培养INH诱导L-02细胞氧化损伤模型,实验分为对照组(A)、INH组(B)、槲皮素低剂量组(C)及槲皮素高剂量组(D)。采用生化分析法检测L-02细胞培养液中天冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)的活性;利用荧光探针检测L-02细胞线粒体内活性氧(ROS)水平;应用比色法检测L-02细胞内丙二醛(MDA)、谷胱甘肽(GSH)的含量以及主要抗氧化物酶的活性。结果表明,与对照组相比,INH能显著增加L-02细胞培养液中AST和ALT的活性、细胞线粒体内ROS水平及细胞内MDA的含量(P<0.01),并显著减少L-02细胞内GSH的含量及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性(P<0.01)。与INH组比较,槲皮素低剂量组L-02细胞培养液中AST的活性、线粒体内ROS水平及细胞内MDA的含量明显降低(P<0.05),而细胞内SOD的活性明显增加(P<0.05);高剂量槲皮素能显著降低L-02细胞培养液中AST和ALT的活性、细胞线粒体内ROS水平及细胞内MDA的含量(P<0.01),并能显著增高L-02细胞内GSH的含量和主要抗氧化物酶的活性(P<0.01)。与槲皮素低剂量组相比,槲皮素高剂量组的保护效应更明显(P<0.05)。可见,ROS介导的氧化应激在INH诱导的L-02细胞毒性中发挥了重要作用,且槲皮素对INH诱导的L-02细胞氧化损伤具有保护作用。
Abstract:
In order to investigate the role of ROS-mediated oxidative stress in INH-induced L-02 cytotoxicity and the intervention of Quercetin, the injury model of hepatocyte L-02 in vitro induced by INH was established. The cells were assigned into the control group (A), INH group (B), Quercetin low dose group (C) and Quercetin high dose group (D). The activity of aspartate aminotransferase (AST) and the activity of alanine transaminase (ALT) in L-02 cells supernatant were determined by biochemical methods; The level of mitochondrial reactive oxy-gen species (ROS) was measured by fluorescent probe 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) in L-02 cells; The content of malondialdehyde (MDA), the content of glutathione (GSH) and the activity of antioxidant enzymes were analyzed with colorimetric method. The results showed that compared with the control group, INH remarkably increased the activity of AST and the activity of ALT in L-02 cells supernatant, the level of mitochondrial ROS and the content of MDA in L-02 cells (P<0.01), and significantly decreased the content of GSH and the activity of superoxide dismutase (SOD), the activity of glutathione peroxidase (GSH-Px) (P<0.01). Compared with the INH group, the activity of AST in L-02 cells supernatant, the level of ROS in mitochondria and the content of MDA in L-02 cells of Quercetin low dose group were decreased (P<0.05), but the activity of SOD was increased (P<0.05); While high dose Quercetin could significantly reduce the activity of AST and the activity of ALT in L-02 cells supernatant, the level of mitochondrial ROS and the content of MDA in L-02 cells (P<0.01), and significantly increased the content of GSH, the activity of SOD and the activity of GSH-Px (P<0.01). Compared with the Quercetin low dose group, the protective effects of high dose Quercetin were more obvious (P<0.05). Therefore, ROS-mediated oxidative stress plays an important role in INH-induced L-02 cell toxicity, and Quercetin has a protective effect on L-02 cell oxidative damage induced by INH.
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