Protective effect of isorhamnetin on oxidative stress injury of HaCaT cells induced by H 2O 2 / 中国医师杂志

ABSTRACT

Objective:To investigate the protective effect of isorhamnetin on oxidative stress injury of HaCaT cells induced by H 2O 2. Methods:HaCaT cells were cultured in vitro and treated with different concentrations of H 2O 2 (300, 600, 900, 1 200 μmol/L) for 12 h. Cell proliferation activity was detected by cell counting kit-8 (CCK-8) assay; SOD activity was detected by superoxide dismutase (SOD) kit and malondialdehyde (MDA) content was detected by MDA assay. The oxidative stress model was established by the selection of suitable H 2O 2 concentration. HaCaT cells were pretreated with isorhamnetin at different concentrations for 12 h, and cell survival rate was detected by CCK-8 method to determine the safe concentration of isorhamnetin for subsequent experiments. HaCaT cells were pretreated with safe concentration of isorhamnetin for 12 h, and H 2O 2 was used to interfere with HaCaT cells for 12 h. Cell proliferation activity, SOD activity and MDA content were detected. Results:With the increase of H 2O 2 concentration, the cell survival rate decreased gradually, the SOD activity decreased gradually and MDA content increased gradually. Compared with the control group, the survival rate of 600, 900 and 1 200 μmol/L H 2O 2 groups was statistically significant ( P<0.05); The SOD activity and MDA content of H 2O 2 groups (300, 600, 900, 1 200 μmol/L) were significantly different from those of the control group ( P<0.05). The oxidative stress model of HaCaT cells was established by 600 μmol/L H 2O 2. HaCaT cells treated with 20, 40, 60, 80 and 100 μmol/L isorhamnetin for 12 h showed no cytotoxic effect. 20, 40 and 60 μmol/L isorhamnetin was selected for subsequent experiments. Compared with H 2O 2 groups, the cell proliferation activity in 40 and 60 μmol/L isornetin groups was significantly increased [(72.21±5.11)%, (76.08±4.91)%, P<0.05], SOD activity increased (19.81±0.38, 20.52±0.52, 15.45±3.13, P<0.05) and MDA content decreased (35.94±0.31, 22.04±0.26, 19.26±1.36, P<0.05). Conclusions:The flavonoid isorhamnetin has a protective effect on oxidative stress injury induced by H 2O 2 in HaCaT cells, suggesting that isorhamnetin may be a potential drug component in the treatment of vitiligo.