Different short- and long-term effects of resveratrol on nuclear factor-kappaB phosphorylation and nuclear appearance in human endothelial cells.
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-kappaB (NF-kappaB)-dependent transcription. Thus, we studied the in vitro influence of physiologic concentrations of resveratrol (<or= 1 micromol/L) on the NF-kappaB signaling pathway after tumor necrosis factor alpha (TNF-alpha) stimulation of endothelial cells.


METHODS
The effects of a 30-min (acute) and an overnight incubation of resveratrol on the nuclear appearance of p50-NF-kappaB and p65-NF-kappaB on serine and tyrosine phosphorylation of the inhibitory subunit kappaB alpha (IkappaBalpha), cytoplasmic concentrations of IkappaBalpha, NF-kappaB phosphorylation or nitrosylation, the reduction of the mitotic inhibitor p21, and the activation of peroxisome proliferator-activated receptor alpha were evaluated.


RESULTS
The nuclear appearance of p50-NFkappaB and p65-NFkappaB acutely induced by TNF-alpha was not modified by resveratrol but was increased after overnight incubation with resveratrol alone or in combination with TNF-alpha. Acute treatment with resveratrol did not modify TNF-alpha-induced cytoplasmic IkappaBalpha serine phosphorylation but did increase IkappaBalpha tyrosine phyophorylation. Resveratrol increased the tyrosine phosphorylation (but not nitrosylation) of immunoprecipitated NF-kappaB, did not decrease cellular p21, and did not increase peroxisome proliferator-activated receptor alpha activity.


CONCLUSIONS
Acute resveratrol treatment does not inhibit the nuclear appearance of NF-kappaB in human umbilical vein endothelial cells, but overnight treatment does. The increase in tyrosine phosphorylation of IkappaBalpha, p50-NF-kappaB, and p65-NF-kappaB suggests the involvement of such alterations in the modulation of NF-kappaB transcription activity.
Pellegatta
Fabio
F
Cardiovascular Pathophysiology Laboratory, Division of Cardiology, Istituto Scientifico Ospedale San Raffaele, Milan, Italy.
Bertelli
Alberto A E
AA
Staels
Bart
B
Duhem
Christian
C
Fulgenzi
Alessandro
A
Ferrero
Maria Elena
ME
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D010766:Phosphorylation / Q000187:drug effects; D000077185:Resveratrol; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013267:Stilbenes / Q000494:pharmacology*; D051996:Transcription Factor RelA; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014443:Tyrosine / Q000378:metabolism
D000894:Anti-Inflammatory Agents, Non-Steroidal; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D013267:Stilbenes; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D014443:Tyrosine; D012694:Serine; D000077185:Resveratrol