Role of Nrf2 signaling pathway in oxidative damage of L-02 cells induced by sodium arsenite / 中华地方病学杂志

ABSTRACT

Objective:To explore the role of nuclear factor-E2-related factor 2 (Nrf2) signaling pathway in oxidative damage caused by sodium arsenite (NaAsO 2) in human normal liver cells (L-02), and to provide experimental basis for the study of oxidative damage mechanism of liver damage caused by arsenic. Methods:L-02 cells were cultured in vitro and treated with 0 (control), 25, 50, 75, 100, 125, and 150 μmol/L NaAsO 2, respectively, for 24 h. The half-inhibitory concentration (IC 50) was calculated according to the cell survival rate by CCK8, and L-02 cells were treated with 0, 1/8, 1/4 and 1/2 dose of IC 50 of NaAsO 2, respectively, for grouping experiments. Protein expressions of Nrf2, heme oxygenase-1 (HO-1), NADH quinone oxidoreductase 1 (NQO1) and glutathione peroxidase 1 (GPx1) in L-02 cells and L-02 nucleus were detected by Western blotting. Results:The result of CCK8 showed that the survival rates of L-02 cells in 25, 50, 75, 100, 125, 150 μmol/L NaAsO 2 groups were [(69.53 ± 0.06)%, (41.33 ± 0.08)%, (23.65 ± 0.04)%, (26.51 ± 0.04)%, (31.63 ± 0.01)%, (26.24 ± 0.02)%], which were significantly lower than that of the control group[(100 ± 0.00)%]. The differences were statistically significant ( P < 0.05). The IC 50 calculated by cell survival was 40 μmol/L, and the NaAsO 2 doses used in the experiment were 0 (control), 5, 10, and 20 μmol/L. Western blotting results showed that, compared with the control group, the protein expression levels of Nrf2, HO-1 in L-02 and HO-1 in the L-02 cells nucleus in the 5, 10 and 20 μmol/L NaAsO 2 groups were significantly higher ( P < 0.05). Compared with the control group, the expression levels of GPx1 protein in L-02 cells of 10 and 20 μmol/L NaAsO 2 groups were decreased ( P < 0.05). Compared with the control group, the expression levels of Nrf2 protein in L-02 nucleus in 10 and 20 μmol/L NaAsO 2 groups were significantly increased ( P < 0.05); the expression level of NQO1 protein in L-02 nucleus in 5 μmol/L NaAsO 2 group was significantly increased ( P < 0.05). Conclusion:NaAsO 2 has an effect on the expression of Nrf2 signaling pathway related factors in L-02 cells, and the mechanism of oxidative damage caused by NaAsO 2 in L-02 cells may be related to Nrf2 signaling pathway.