Red wine polyphenols increase calcium in bovine aortic endothelial cells: a basis to elucidate signalling pathways leading to nitric oxide production

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. 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. RWPC, Provinols™ and delphinidin increased cytosolic free calcium ([Ca 2+ ] i ), by releasing Ca 2+ from intracellular stores and by increasing Ca 2+ entry. The RWPC‐induced increase in [Ca 2+ ] i was decreased by exposure to ryanodine (30 μ M ), whereas Provinols™ and delphinidin‐induced increases in [Ca 2+ ] i were decreased by bradykinin (0.1 μ M ) and thapsigargin (1 μ M ) pre‐treatment. RWPC, Provinols™ and delphinidin‐induced increases in [Ca 2+ ] i were sensitive to inhibitors of phospholipase C (neomycin, 3 m M ; U73122, 3 μ M ) and tyrosine kinase (herbimycin A, 1 μ M ). RWPC, Provinols™ and delphinidin induced herbimycin A (1 μ M )‐sensitive tyrosine phosphorylation of several intracellular proteins. 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. 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.British Journal of Pharmacology (2002) 135 , 1579–1587; doi: 10.1038/sj.bjp.0704603