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目的探讨五价砷酸钠暴露对Chang肝细胞核转录因子-E2类相关因子2(Nrf2)及其调控的下游抗氧化酶NAD(P)H:醌氧化还原酶1(NQO1)和血红素单加氧酶-1(HO-1)的mRNA及其蛋白表达的影响。方法 0.5、1.0、2.0mmol/L的砷酸钠(Na2HAsO4)染毒Chang肝细胞6 h后,采用western blot法检测细胞内Nrf2、NQO1和HO-1的蛋白表达情况;采用抗氧化反应元件(ARE)-荧光素酶活性检测Nrf2的转录活性情况;采用Real-time PCR检测细胞NQO1和HO-1的mRNA表达情况。结果 0.5、1.0、2.0 mmol/L的Na2HAsO4染毒6 h均能够显著诱导Chang细胞的Nrf2蛋白表达(P<0.05),ARE荧光素酶活性水平也随染毒剂量的增加而诱导,并且具有剂量-效应关系;随着染毒剂量的升高,HO-1和NQO1的mRNA及蛋白水平逐渐升高,与对照组相比具有统计学差异,并且具有剂量-效应关系(P<0.01)。结论砷酸钠暴露能够诱导人类Chang肝细胞的Nrf2及其调控的下游抗氧化酶NQO1和HO-1的mRNA及其蛋白的表达。
Objective To investigate the effects of pentavalent arsenite exposure on the expression of Nrf2 and its downstream NAD (P) H: quinone oxidoreductase 1 (NQO1) and heme alone in Chang hepatocytes. Oxygenase-1 (HO-1) mRNA and Its Protein Expression. Methods The hepatocytes were treated with 0.5,1.0,2.0 mmol / L sodium arsenate (Na2HAsO4) for 6 h, and the protein expression of Nrf2, NQO1 and HO-1 were detected by western blot. Antioxidant elements ARE) - luciferase activity assay Nrf2 transcriptional activity; Real-time PCR detection of cell NQO1 and HO-1 mRNA expression. Results Nrf2 protein expression in Chang cells was significantly induced by 0.5, 1.0, 2.0 mmol / L Na2HAsO4 for 6 h (P <0.05), and the concentration of ARE luciferase was also induced with an increase in the dose of Na2HAsO4. -effect relationship. The mRNA and protein levels of HO-1 and NQO1 gradually increased with the increase of exposure dose, which were statistically different from the control group and had a dose-effect relationship (P <0.01). Conclusion Sodium arsenate exposure can induce the expression of Nrf2 and its downstream antioxidant enzymes NQO1 and HO-1 mRNA and protein in human Chang hepatocytes.