RESUMO
We examined the relaxant effects of natriuretic peptide family on the isolated guinea-pig aorta to determine the receptor subtype which primarily mediates this vascular relaxation, with particular attention to the apparent contribution of voltage-dependent and Ca2+-activated KS (BK(Ca)) channels to the response. Three endogenous natriuretic peptide ligands (natriuretic peptide, ANP; brain natriuretic peptide, BNP; C-type natriuretic peptide, CNP) produced a concentration-dependent relaxation in de-endothelialized guinea-pig aorta pre-contracted by noradrenaline (NA), with a potency order of ANP > or = BNP >> CNP. Although the relaxations elicited by these three natriuretic peptide ligands were significantly diminished by iberiotoxin (IbTx, 10(-7) M), a selective BK(Ca) channel blocker, the inhibitory effect of IbTx was most pronounced for the CNP-induced relaxation; when estimated at 10(-7) M of each peptide, the apparent extent of BK(Ca) channel contribution to the total relaxant response was approximately 60% for CNP > approximately 20% for either ANP or BNP. Supporting the substantial role of BK(Ca) channels in the vascular responses, high-KCl (80 mM) potently suppressed the relaxations induced by these natriuretic peptide ligands. The relaxant response to 8-Bromo-cyclic GMP, a membrane permeable cyclic GMP analogue, was also diminished by IbTx (10(-7) M) and high-KCl (80 mM), which indicates the key role of cyclic GMP in the BK(Ca) channel-mediated, natriuretic peptide-elicited vascular relaxation. These results indicate that the A-type receptor (NPR-A, which is more selective for ANP and BNP) rather than the B-type receptor (NPR-B, which is more selective for CNP) predominates in the guinea-pig aorta as the natriuretic peptide receptor which mediates this vascular smooth muscle relaxation. Although activation of BK(Ca) channels substantially contributes to both NPR-A- and NPR-B-activated relaxations, particularly in the NPR-B-activated relaxation, this K channel may function as a primary relaxant mediator in this conduit artery.
Assuntos
Aorta Torácica/efeitos dos fármacos , Aorta Torácica/fisiologia , Fator Natriurético Atrial/farmacologia , GMP Cíclico/análogos & derivados , Peptídeo Natriurético Encefálico/farmacologia , Peptídeo Natriurético Tipo C/farmacologia , Vasodilatação , Animais , GMP Cíclico/farmacologia , Feminino , Cobaias , Técnicas In Vitro , Canais de Potássio Ativados por Cálcio de Condutância Alta , Masculino , Músculo Liso Vascular , Canais de Potássio Cálcio-Ativados/fisiologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/fisiologia , Cloreto de Potássio/administração & dosagem , Vasoconstrição/efeitos dos fármacos , Vasodilatação/fisiologiaRESUMO
The role of K(+) channels in nitric oxide (NO)-induced vasorelaxation has been largely investigated in resistance vessels where iberiotoxin-sensitive MaxiK channels play a predominant role. However, the nature of the K(+) channel(s) involved in the relaxation triggered by NO-releasing compounds [nitroglycerin, NTG; NOR 3 [(+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide]] or atrial natriuretic peptide (ANP) in the conduit vessel aorta has remained elusive. We now demonstrate that, in rat aorta, the relaxation due to these vasorelaxants is not affected by the MaxiK channel blocker iberiotoxin (10(-7)-10(-6) M) as was the control vascular bed used (mesenteric artery). The inability of iberiotoxin to prevent NO/ANP-induced aortic relaxations was not due to lower expression of MaxiK in aorta or due to the predominance of iberiotoxin-resistant channels in this conduit vessel. Aortic relaxations were strongly diminished by 4-aminopyridine (4-AP) (> or =5 x 10(-3) M) or by tetraethylammonium (>2 x 10(-3) M) at concentrations known to inhibit voltage-dependent K(+) (K(v)) 2-type channels but not by other K(+) channel inhibitors, glibenclamide, apamin, charybdotoxin, tertiapin, or E-4031 N-[4-[[1-[2-(6-methyl-2-pyridinyl)ethyl]-4-piperidinyl-]carbonyl]phenyl]methanesulfonamide dihydrochloride). Consistent with a role of K(v)2-type channels, K(v) currents in A7r5 aortic myocytes were stimulated by NTG and inhibited by > or =5 x 10(-3) M 4-AP. Furthermore, immunocytochemistry, immunoblot, and real-time polymerase chain reaction analyses confirmed the presence of K(v)2.1 channels in aorta. K(v)2.1 transcripts were approximately 100-fold more abundant than K(v)2.2. Our results support low-affinity 4-AP-sensitive K(v) channels, assembled at least partially by K(v)2.1 subunit, as downstream effectors of NO/ANP-signaling cascade regulating aortic vasorelaxation and further demonstrate vessel-specific K(+) channel involvement in NO/ANP-induced relaxation.
Assuntos
Aorta Torácica/fisiologia , Fator Natriurético Atrial/fisiologia , Artérias Mesentéricas/fisiologia , Óxido Nítrico/fisiologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/fisiologia , Vasodilatação/fisiologia , Animais , Aorta Torácica/efeitos dos fármacos , Aorta Torácica/metabolismo , Fator Natriurético Atrial/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Membrana Celular/fisiologia , Inibidores Enzimáticos/farmacologia , Guanilato Ciclase , Immunoblotting , Técnicas In Vitro , Canais de Potássio Ativados por Cálcio de Condutância Alta/biossíntese , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/metabolismo , Óxido Nítrico/metabolismo , Doadores de Óxido Nítrico/farmacologia , Bloqueadores dos Canais de Potássio/farmacologia , Canais de Potássio de Abertura Dependente da Tensão da Membrana/antagonistas & inibidores , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Guanilil Ciclase Solúvel , Vasodilatação/efeitos dos fármacosRESUMO
Inhibitory effects of docosahexaenoic acid (DHA) on the muscle contractions induced by U46619, a thromboxane A2 (TXA2) mimetic, and phenylephrine were compared in guinea-pig aorta. In de-endothelialized guinea-pig aortic ring preparations, DHA at 10 microM strongly inhibited a sustained contraction produced by U46619 (3-100 nM) whereas it did not exhibit an appreciable effect on phenylephrine (3-10 microM)-induced contraction. The present findings indicate that DHA inhibits more selectively TXA2 receptor (TP receptor)-mediated vascular contraction than alpha-adrenoceptor-mediated response. Selective inhibition by DHA of TP receptor-mediated contraction of blood vessels seems underlie in part the mechanisms by which this polyunsaturated fatty acid exerts its circulatory-protective effects.