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目的探讨富勒烯(C60)对人胚肝细胞(L-02)内过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)mRNA表达的影响。方法将对数生长期的人胚肝细胞接种于96孔培养板(细胞密度为104个/孔),随机分为对照组(生理盐水)和1.25、2.50、5.00、10.00、20.00、40.00μg/ml富勒烯染毒组,培养24h后,采用MTT试验检测富勒烯对细胞存活率的影响。将人胚肝细胞分别暴露于0、1.00和20.00μg/ml富勒烯24h,采用实时定量PCR法检测细胞内CAT、GSH-Px和SODmRNA的相对表达量。结果与对照组比较,1.25、2.50、5.00、10.00、20.00、40.00μg/ml富勒烯染毒组的细胞存活率均降低,差异有统计学意义(P<0.05或P<0.01);且细胞存活率随富勒烯染毒剂量的增加而降低。与对照组比较,1.00μg/ml富勒烯染毒组GSH-Px和SOD以及20.00μg/ml富勒烯染毒组CAT、GSH-Px和SODmRNA相对表达水平较低,差异均有统计学意义(P<0.05或P<0.01);且人胚肝细胞中CAT、GSH-Px、SODmRNA的相对表达水平随着富勒烯染毒剂量的增加而降低。结论富勒烯可通过降低人胚胎肝细胞内CAT、GSH-Px和SODmRNA的表达水平,引起肝细胞发生过氧化损伤。
Objective To investigate the effects of fullerene (C60) on catalase (CAT), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) mRNA in human embryonic hepatocytes (L-02) The impact of expression. Methods Human embryonic hepatocytes in logarithmic growth phase were inoculated into 96-well plates (cell density 104 cells / well) and randomly divided into control group (normal saline) and 1.25, 2.50, 5.00, 10.00, 20.00 and 40.00 μg / ml fullerene. After cultured for 24 h, the effect of fullerene on cell viability was tested by MTT assay. The human embryonic hepatocytes were exposed to 0, 1.00 and 20.00μg / ml fullerenes respectively for 24 hours. The relative expression levels of CAT, GSH-Px and SODmRNA were detected by real-time quantitative PCR. Results Compared with the control group, the cell viability in the groups treated with fulleretin at 1.25, 2.50, 5.00, 10.00, 20.00 and 40.00 μg / ml was significantly lower than that in the control group (P <0.05 or P <0.01) Survival rate decreased with increasing doses of fullerenes. Compared with the control group, the relative expression levels of GSH-Px and SOD and the CAT, GSH-Px and SODmRNA in 1.00μg / ml Fullerene-exposed group and 20.00μg / ml fullerene-exposed group were lower, the differences were statistically significant (P <0.05 or P <0.01). The relative expression levels of CAT, GSH-Px and SODmRNA in human embryonic hepatocytes decreased with the increase of fullerene dose. Conclusion Fullerenes can induce the peroxidation injury of hepatocytes by reducing the expression of CAT, GSH-Px and SODmRNA in human embryonic liver cells.