Effects of resveratrol on endothelial nitric oxide synthase (eNOS) phosphorylation and interaction with regulatory proteins

Previous studies have demonstrated that the polyphenolic compound, reseveratrol (RSV), from the skin of red grapes increases endothelial nitric oxide synthase (eNOS) activity. However, the mechanism by which RSV increases eNOS activity is unknown. Therefore, we have investigated changes in eNOS phosphorylation at Ser1179, Ser635, Thr495 as well as changes in eNOS association with Hsp90, Akt, and cdc37 in bovine aortic endothelial cells (BAECs) treated with varying doses of RSV for 0, 20, 60 min as well as 72 h. Acute RSV treatment (less than 1) resulted in no significant ( P >0.05) changes in eNOS phosphorylation or changes in eNOS association with Hsp90, Akt or cdc37. In contrast, 72h treatment of BAECs at higher concentrations of RSV significantly ( P <0.05) increased eNOS phosphorylation and protein association that was consistent with the increase in eNOS expression. In conclusion, these data suggest that RSV‐induced eNOS activity is primarily due to increased eNOS expression and that acute (less than 1 h) RSV treatment does not alter post‐translational eNOS regulatory mechanisms. Supported by AHA SDG 0430157N (MBH) and William & Mary HHMI Undergraduate Research Program Student Grant (KNS)