Resveratrol, a polyphenolic phytoalexin present in red wine, enhances expression and activity of endothelial nitric oxide synthase.
BACKGROUND
Estrogens can upregulate endothelial nitric oxide synthase (eNOS) in human endothelial cells by increasing eNOS promoter activity and enhancing the binding activity of the transcription factor Sp1. Resveratrol, a polyphenolic phytoalexin found in grapes and wine, has been reported to act as an agonist at the estrogen receptor. Therefore, we tested the effect of this putative phytoestrogen on eNOS expression in human endothelial cells.


RESULTS
Incubation of human umbilical vein endothelial cells (HUVEC) and HUVEC-derived EA.hy 926 cells with resveratrol for 24 to 72 hours upregulated eNOS mRNA expression in a time- and concentration-dependent manner (up to 2.8-fold). eNOS protein expression and eNOS-derived NO production were also increased after long-term incubation with resveratrol. Resveratrol increased the activity of the eNOS promoter (3.5-kb fragment) in a concentration-dependent fashion, with the essential trans-stimulated sequence being located in the proximal 263 bp of the promoter sequence. In addition, eNOS mRNA was stabilized by resveratrol. The effect of resveratrol on eNOS expression was not modified by the estrogen receptor antagonists ICI 182780 and RU 58668. In electrophoretic mobility shift assays, nuclear extracts from resveratrol-incubated EA.hy 926 cells showed no enhanced binding activity of the eNOS promoter-relevant transcription factors Sp1, GATA, PEA3, YY1, or Elf-1. In addition to its long-term effects on eNOS expression, resveratrol also enhanced the production of bioactive NO in the short-term (after a 2-minute incubation).


CONCLUSIONS
In concert with other effects, the stimulation of eNOS expression and activity may contribute to the cardiovascular protective effects attributed to resveratrol.
Wallerath
Thomas
T
Department of Pharmacology, Johannes Gutenberg University, Mainz, Germany.
Deckert
Göran
G
Ternes
Thomas
T
Anderson
Henrik
H
Li
Huige
H
Witte
Klaus
K
Förstermann
Ulrich
U
D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D020536:Enzyme Activators / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D004965:Estrogen Antagonists / Q000494:pharmacology; D004968:Estrogens, Non-Steroidal / Q000032:analysis / Q000494:pharmacology; D005419:Flavonoids; D006801:Humans; D007529:Isoflavones; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000235:genetics / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III; D023282:Nuclease Protection Assays; D010636:Phenols / Q000494:pharmacology*; D048789:Phytoestrogens; D010936:Plant Extracts / Q000494:pharmacology*; D028321:Plant Preparations; D011108:Polymers / Q000494:pharmacology*; D059808:Polyphenols; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000502:physiology; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors; D000077185:Resveratrol; D012717:Sesquiterpenes; D013267:Stilbenes / Q000494:pharmacology*; D013729:Terpenes; D015854:Up-Regulation / Q000187:drug effects; D014920:Wine / Q000032:analysis; D000094162:Phytoalexins
D020536:Enzyme Activators; D004965:Estrogen Antagonists; D004968:Estrogens, Non-Steroidal; D005419:Flavonoids; D007529:Isoflavones; D010636:Phenols; D048789:Phytoestrogens; D010936:Plant Extracts; D028321:Plant Preparations; D011108:Polymers; D059808:Polyphenols; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D012717:Sesquiterpenes; D013267:Stilbenes; D013729:Terpenes; D009569:Nitric Oxide; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D000077185:Resveratrol; D000094162:Phytoalexins