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Daidzein Has Anti-Oxidant Activity in Normal Human Kidney Tubular HK-2 Cells via FOXO3/SOD2 Pathway

Jongsung Lee, See-Hyoung Park

Journal of Food and Nutrition Research. 2018, 6(9), 557-560 doi:10.12691/jfnr-6-9-3
  • Figure 1. Daidzein has anti-oxidant activity through up-regulation of SOD2 expression in HK-2 cells. A) Decreased ROS level in HK-2 cells treated with daidzein. ROS level in HK-2 cells was measured by ROS detection kit B) Treatment of daidzein in HK-2 cells caused up-regulation of SOD2 expression with dose-dependency. Western blotting analysis of SOD1, SOD2, and SOD3 expression in HK-2 cells. C) Treatment of daidzein in HK-2 cells caused up-regulation of SOD2 expression with time-dependency. Western blotting analysis of SOD1, SOD2, and SOD3 expression in HK-2 cells. β-actin was used for loading control. D) Dose-dependent WST-1 assay showed that daidzein has no cytotoxicity on HK-2 cells. E) Cell counting assay was performed to check daidzein treatment in HK-2 cells affect the cell growth compared to DMSO control for 0, 24, 48, and 72 h. Significant between time group are indicated (* means P < 0.05). , error bars represent standard deviation, and the statistical test paired t-test
  • Figure 2. Involvement of FOXO3 to mediate SOD2 expression by daidzein. A) Protein from HK-2 cell treated with daidzein was prepared by the cytosol and nucleous fractional method described in material and method section. Then, FOXO3 and SOD2 expression level was analyzed by Western blotting. β-actin was used for cytosol loading control and PARP1 was for nucleous loading control. B) Immunofluorescence analysis of FOXO3 in HK-2 cells treated with daidzein was performed by the method described in the methods. C) siRNA against FOXO3 was transfected into HK-2 cells followed by daidzein treatment. Then, FOXO3 and SOD2 expression level was analyzed by Western blotting. β-actin was used for loading control D) siRNA against FOXO3 was transfected into HK-2 cells followed by daidzein treatment. Then, ROS level in HK-2 cells was measured by ROS detection kit