Red wine polyphenols increase calcium in bovine aortic endothelial cells: a basis to elucidate signalling pathways leading to nitric oxide production.
1. The present study investigates the mechanisms by which polyphenolic compounds from red wine elicit Ca(2+) mobilization in bovine aortic endothelial cells (BAECs). Two polyphenol-containing red wine extracts, red wine polyphenolic compounds (RWPC) and Provinols, and delphinidin, an anthocyanin were used. 2. RWPC stimulated a Ca(2+)-dependent release of nitric oxide (NO) from BAECs accounting for the relaxation of endothelium-denuded rat aortic rings as shown by cascade bioassay. 3. RWPC, Provinols and delphinidin increased cytosolic free calcium ([Ca(2+)](i)), by releasing Ca(2+) from intracellular stores and by increasing Ca(2+) entry. 4. The RWPC-induced increase in [Ca(2+)](i) was decreased by exposure to ryanodine (30 microM), whereas Provinols and delphinidin-induced increases in [Ca(2+)](i) were decreased by bradykinin (0.1 microM) and thapsigargin (1 microM) pre-treatment. 5. RWPC, Provinols and delphinidin-induced increases in [Ca(2+)](i) were sensitive to inhibitors of phospholipase C (neomycin, 3 mM; U73122, 3 microM) and tyrosine kinase (herbimycin A, 1 microM). 6. RWPC, Provinols and delphinidin induced herbimycin A (1 microM)-sensitive tyrosine phosphorylation of several intracellular proteins. 7. Provinols released Ca(2+) via both a cholera (CTX) and pertussis toxins (PTX)-sensitive pathway, whereas delphinidin released Ca(2+) only via a PTX-sensitive mechanism. 8. Our data contribute in defining the mechanisms of endothelial NO production caused by wine polyphenols including the increase in [Ca(2+)](i) and the activation of tyrosine kinases. Furthermore, RWPC, Provinols and delphinidin display differences in the process leading to [Ca(2+)](i) increases in endothelial cells illustrating multiple cellular targets of natural dietary polyphenolic compounds.
Martin
Sophie
S
Pharmacologie et Physico-Chimie des Interactions Cellulaires et Moléculaires, UMR CNRS 7034, Faculté de Pharmacie, Université Louis Pasteur, 67401 Illkirch, France.
Andriambeloson
Emile
E
Takeda
Ken
K
Andriantsitohaina
Ramaroson
R
D000818:Animals; D000872:Anthocyanins; D001011:Aorta / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D001578:Benzopyrans / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism* / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D005110:Extracellular Space / Q000378:metabolism / Q000502:physiology; D005419:Flavonoids; D009569:Nitric Oxide / Q000096:biosynthesis*; D010636:Phenols / Q000494:pharmacology*; D011108:Polymers / Q000494:pharmacology*; D059808:Polyphenols; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D010738:Type C Phospholipases / Q000502:physiology; D014664:Vasodilation / Q000187:drug effects / Q000502:physiology; D014920:Wine
D000872:Anthocyanins; D001578:Benzopyrans; D005419:Flavonoids; D010636:Phenols; D011108:Polymers; D059808:Polyphenols; D009569:Nitric Oxide; D002772:Cholera Toxin; D011505:Protein-Tyrosine Kinases; D010738:Type C Phospholipases; C017185:delphinidin; D002118:Calcium