title: Protein kinase C activation increases endothelial nitric oxide release in mesenteric arteries from orchidectomized rats.
abstract: The aim of the present study was to assess the effect of endogenous male sex hormones on endothelial nitric oxide synthase (eNOS) expression, release and function of the endothelial nitric oxide (NO), as well as to assess the regulatory action of protein kinase C (PKC) on acetylcholine (ACh)-induced endothelial NO release. For this purpose, superior mesenteric arteries from control and orchidectomized male Sprague-Dawley rats were used. eNOS expression and basal-and ACh-induced NO release were similar in arteries from both groups of rats. Orchidectomy decreased the vasodilator effect induced by ACh but did not alter that induced by sodium nitroprusside (SNP). The superoxide anion scavenger, superoxide dismutase (SOD), or the membrane-permeable mimetic of SOD, tempol, only enhanced ACh-induced relaxation in arteries from orchidectomized rats. ACh-induced TXA(2) formation was higher in arteries from orchidectomized than from control rats. Neither the PKC activator, phorbol 12,13-dibutyrate (PDBu), nor the non-selective PKC inhibitor, calphostin C, modified basal- or ACh-induced NO release in arteries from control rats. In arteries from orchidectomized rats, basal- and ACh-induced endothelial NO release were increased by PDBu but decreased by calphostin C. Both Gö6976, a PKC inhibitor that is partially selective for conventional PKC isoforms, as well as PKCzeta pseudosubstrate inhibitor (PKCzeta-PI) decreased both basal- and ACh-induced NO release in arteries from orchidectomized rats. Neither PDBu nor calphostin C modified the vasodilator response induced by ACh in arteries from control rats. In segments from orchidectomized rats, PDBu enhanced the ACh-induced response, but this response was not modified by calphostin C, Gö6976 or PKCzeta-PI. The vasodilator response induced by SNP was not altered by the PKC activators or inhibitors in any artery from either group. These results show that endogenous male sex hormone deprivation does not affect the eNOS expression or the endothelial NO release induced by ACh, but does decrease the vasodilator action of ACh, by increasing NO metabolism and TXA(2) formation. In addition, PKC seems to modulate eNOS activity only in mesenteric arteries from orchidectomized rats, in which conventional and PKCzeta isoforms are involved in the positive regulation of eNOS.
authors.affiliation: Departamento de Fisiología, Facultad de Medicina, Universidad Autónoma de Madrid, C/Arzobispo Morcillo 4, 28029 Madrid, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Blanco-Rivero
authors.forename: Ana
authors.initials: A
authors.lastname: Sagredo
authors.forename: Gloria
authors.initials: G
authors.lastname: Balfagón
authors.forename: Mercedes
authors.initials: M
authors.lastname: Ferrer
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002227:Carbazoles / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008638:Mesenteric Arteries / Q000201:enzymology*; D009281:Naphthalenes / Q000494:pharmacology; D009569:Nitric Oxide / Q000032:analysis; D052250:Nitric Oxide Synthase Type III / Q000032:analysis / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology; D009919:Orchiectomy; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013268:Stimulation, Chemical; D013481:Superoxides / Q000378:metabolism; D013928:Thromboxane A2 / Q000032:analysis / Q000378:metabolism; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D002227:Carbazoles; D007211:Indoles; D009281:Naphthalenes; D014665:Vasodilator Agents; D013481:Superoxides; C081021:Go 6976; D009599:Nitroprusside; D009569:Nitric Oxide; D015240:Phorbol 12,13-Dibutyrate; D013928:Thromboxane A2; D052250:Nitric Oxide Synthase Type III; C087857:protein kinase C gamma; D011493:Protein Kinase C; C058819:calphostin C; D000109:Acetylcholine