Effects and mechanisms of resveratrol on hydrogen peroxide-induced human arterial smooth muscle cells

Objective:To investigate the effects and mechanisms of resveratrol(Res) on hydrogen peroxide(H_2O_2)-induced human arterial smooth muscle cells.Methods;Human arterial smooth muscle cells were incubated with H_2O_2(20,100 and 500|xmol/L) for 24h to establish the oxidative stress-induced cell injury model.This was followed by the addition of different concentrations of Res(2,10 and 50μmol/L) with or without H_2O_2(l00μmol/L) to the arterial smooth muscle cells for 24h.The viability of human arterial smooth muscle cells was measured by MTT assay,and the SOD activity was assayed by using xanthine oxidase method.Thiobarbituric acid colorimetric approach was employed to determine the level of MDA.The level of reactive oxygen species(ROS) was detected by DCFH-DA staining and photofluorography,and the percentage of apoptotic cells was assessed by using flow cytometry(FCM).Results:Following treatment with 20,100 and 500 μmol/L H_2O_2 for 24h,there was a reduction in the cell viability and SOD activity in the supernatant whilst increased level of MDA,in a concentration-dependent manner,in the human arterial smooth muscle cell supernatant(all P 0.05).Incubation with 2,10 and 50μmol/L Res and 100μmol/L H_2O_2 for 24 h ameliorated the reduced cell vitality,SOD activity and the level of MDA in a dose-dependent fashion.The differences in SOD activity differed substantially between 100 μmol/L H_2O_2 model group and 10 and 50 μmol/L Res plus 100μmol/L H_2O_2 treatment groups(both P0.05).Compared with 100 μmol/L H_2O_2 model group,the DCF fluorescence intensity was significantly increased in 10 and 50μmol/L Res plus 100 μmol/L H_2O_2 treatment groups(both P 0.05).The 10 and50μmol/L plus 100μmol/L H_2O_2 treatment groups were associated with reduced apoptosis rate compared with100 μmol/LH_2O_2 model group[(20.7±3.0)%and(13.2 ± 1.5)%vs.(27.2 ±3.6)%].Conclusion;Res protected human arterial smooth muscle cells against H_2O_2-induced oxidative stress injury,presumably associated with reduced production of Res and apoptosis rate of vascular smooth muscle cells.