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Original Research Communication |
B phosphorylation and nuclear appearance in human endothelial cells1,2
1 From the Cardiovascular Pathophysiology Laboratory, Division of Cardiology, Istituto Scientifico Ospedale San Raffaele, Milan, Italy (FP); the Department of Human Anatomy, University of Milan, Milan, Italy (AAEB); INSERM, Institut Pasteur de Lille, Lille, France (BS and CD); and Istituto di Patologia Generale, Universitá degli Studi di Milano, Milan, Italy (AF and MEF).
Background: Resveratrol (a naturally occurring phytoalexin found in grapes and wine) has cardiovascular protective effects that suggest the antiatherogenic (ie, antiinflammatory) activities of the compound on endothelial cells.
Objective: The antiinflammatory activity of resveratrol could be mediated by its interference with nuclear factor-
B (NF-B)–dependent transcription. Thus, we studied the in vitro influence of physiologic concentrations of resveratrol (
1 µmol/L) on the NF-B signaling pathway after tumor necrosis factor 
(TNF-) stimulation of endothelial cells.
Design: The effects of a 30-min (acute) and an overnight incubation of resveratrol on the nuclear appearance of p50-NF-B and p65-NF-B on serine and tyrosine phosphorylation of the inhibitory subunit B (IB), cytoplasmic concentrations of IB, NF-B phosphorylation or nitrosylation, the reduction of the mitotic inhibitor p21, and the activation of peroxisome proliferator–activated receptor were evaluated.
Results: The nuclear appearance of p50-NFB and p65-NFB acutely induced by TNF- was not modified by resveratrol but was increased after overnight incubation with resveratrol alone or in combination with TNF-. Acute treatment with resveratrol did not modify TNF-–induced cytoplasmic IB serine phosphorylation but did increase IB tyrosine phyophorylation. Resveratrol increased the tyrosine phosphorylation (but not nitrosylation) of immunoprecipitated NF-B, did not decrease cellular p21, and did not increase peroxisome proliferator–activated receptor activity.
Conclusions: Acute resveratrol treatment does not inhibit the nuclear appearance of NF-B in human umbilical vein endothelial cells, but overnight treatment does. The increase in tyrosine phosphorylation of IB, p50-NF-B, and p65-NF-B suggests the involvement of such alterations in the modulation of NF-B transcription activity.
Key Words: Resveratrol • endothelial cells • nuclear factor-B • atherosclerosis • inflammatory disease • red wine • tumor necrosis factor
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