Betaine Increasing Expression Levels of Antioxidase in LO2 Cells
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Abstract:
The present study investigated the effect of betaine (BET) on the mRNA levels of superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) in human normal hepatocyte LO2 cells and the mechanism. The mRNA levels of antioxidases and indicators involving Nrf2-Keap1-ARE pathway were detected by real-time fluorescence quantitative method, western blot and enzyme-linked immunosorbent assay. The mRNA levels of SOD, GPX and CAT in LO2 cells were significantly increased by 1.08, 0.51 and 0.88 times, respectively, after 12 h treatment with different concentrations of BET (p<0.05). After 250 μmol/L of BET treatment, the protein level of Nrf2, which can bind to the ARE region in the nucleus, and the total Nrf2 protein in cells were significantly increased by 1.21 and 0.80 fold, respectively (p<0.05).After treatment with brucopicol, the effect of BET on the mRNA levels of three antioxidant enzymes was completely abolished. Further analysis showed that BET did not significantly increase the mRNA and phosphorylation levels of Nrf2, but decreased the mRNA and protein content of Keap1 by 38.21% and 49.15%, respectively (p<0.05). These results indicate that BET activates the Nrf2-Keap1-ARE pathway by decreasing the level of Keap1, and thus enhances the expression of antioxidant enzymes.