Original Article
Role of Nrf2 signaling pathway in oxidative damage of L-02 cells induced by sodium arsenite
Li Xiaozhi, Hu Ting, Duan Tianxiao, Zhang Qi, Wu Changyan, Jian Wen, Luo Peng
Published 2020-04-20
Cite as Chin J Endemiol, 2020, 39(4): 259-263. DOI: 10.3760/cma.j.cn231583-20191104-00308
Abstract
ObjectiveTo explore the role of nuclear factor-E2-related factor 2 (Nrf2) signaling pathway in oxidative damage caused by sodium arsenite (NaAsO2) 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.
MethodsL-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 (IC50) 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 IC50 of NaAsO2, 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.
ResultsThe 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).
ConclusionNaAsO2 has an effect on the expression of Nrf2 signaling pathway related factors in L-02 cells, and the mechanism of oxidative damage caused by NaAsO2 in L-02 cells may be related to Nrf2 signaling pathway.
Key words:
Arsenites; Liver cells; Nrf2 signaling pathway; Oxidative damage
Contributor Information
Li Xiaozhi
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Hu Ting
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Key Laboratory for Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, China
Duan Tianxiao
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Zhang Qi
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Wu Changyan
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Jian Wen
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Luo Peng
School of Public Health, Guizhou Medical University, Guiyang 550025, China
Key Laboratory for Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 550025, China
