title: Signaling pathways involved in the H2O2-induced vasoconstriction of rat coronary arteries.
abstract: Hydrogen peroxide (H2O2) is an endogenous endothelium-derived hyperpolarizing factor released by flow and involved in the regulation of coronary blood flow. Because opposing vasoactive effects have been reported for H2O2 depending on the vascular bed and experimental conditions, the aim of this study was to assess whether H2O2 may act as a coronary vasoconstrictor and if so to determine the underlying signaling mechanisms. Intramyocardial arteries from male Wistar rats were mounted on microvascular myographs for simultaneous measurements of intracellular Ca(2+) ([Ca(2+)]i) and tension. On coronary arteries precontracted with the thromboxane A2 (TxA2) analogue U46619, H2O2 (1-300μM) elicited further moderate contractions in the proximal arterial segments and relaxed the more distal coronary branches, the contractions being markedly augmented in arteries depolarized by raising extracellular K(+). H2O2-elicited vasoconstriction on K(+)30-precontracted coronary arteries was blunted by catalase and significantly reduced by endothelial cell removal and by inhibitors of cyclooxygenase (COX) and of the TxA2 receptor (TP). H2O2 (50μM) increased by about 10-fold basal superoxide anion (O2(-)) production in coronary arteries measured by lucigenin-enhanced chemiluminescence, and H2O2-elicited contractions were reduced by the superoxide dismutase mimetic tempol and by NADPH oxidase inhibition. Furthermore, blockade of the ERK and p38 mitogen-activated protein (MAP) kinases significantly reduced the contractions elicited by high and low concentrations of peroxide, respectively, whereas Rho kinase inhibition nearly abolished these responses. H2O2 (50μM) elicited simultaneous and similar sustained increases in [Ca(2+)]i and tension that were blunted by blockade of voltage-dependent L-type channels, but resistant to the nonselective Ca(2+) channel blocker 2-aminoethoxydiphenyl borate. Moreover, endothelial cell removal reduced the increases in [Ca(2+)]i and contraction elicited by peroxide. The present data demonstrate that H2O2 is an endothelium-dependent vasoconstrictor in rat coronary arteries that activates smooth muscle Ca(2+) entry through L-type and non-L-type channels and various intracellular signaling pathways including the release of a COX-derived TP agonist, stimulation of the MAP and Rho kinase pathways, and production of NADPH oxidase-derived superoxide.
authors.affiliation: Departamento de Fisiología, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain.
authors.forename: Elvira
authors.initials: E
authors.lastname: Santiago
authors.forename: Cristina
authors.initials: C
authors.lastname: Contreras
authors.forename: Albino
authors.initials: A
authors.lastname: García-Sacristán
authors.forename: Ana
authors.initials: A
authors.lastname: Sánchez
authors.forename: Luis
authors.initials: L
authors.lastname: Rivera
authors.forename: Belén
authors.initials: B
authors.lastname: Climent
authors.forename: Dolores
authors.initials: D
authors.lastname: Prieto
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism; D003331:Coronary Vessels / Q000187:drug effects / Q000378:metabolism*; D004727:Endothelium / Q000187:drug effects / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000008:administration & dosage*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D051381:Rats; D044244:Receptors, Thromboxane A2, Prostaglandin H2 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013481:Superoxides / Q000378:metabolism / Q000494:pharmacology; D013928:Thromboxane A2 / Q000494:pharmacology; D014661:Vasoconstriction; D014662:Vasoconstrictor Agents / Q000008:administration & dosage
chemical_list: D020746:Calcium Channels, L-Type; D044244:Receptors, Thromboxane A2, Prostaglandin H2; D014662:Vasoconstrictor Agents; D013481:Superoxides; D013928:Thromboxane A2; D006861:Hydrogen Peroxide; D011451:Prostaglandin-Endoperoxide Synthases; D002118:Calcium