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Id: lil-722165
Autor: Leite, L.N.; Gonzaga, N.A.; Tirapelli, D.P.C.; Tirapelli, L.F.; Tirapelli, C.R..
Título: Pharmacological characterization of the relaxant effect induced by adrenomedullin in rat cavernosal smooth muscle
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;47(10):876-885, 10/2014. tab, graf.
Idioma: en.
Projeto: FAPESP.
Resumo: The aim of the present study was to determine the mechanisms underlying the relaxant effect of adrenomedullin (AM) in rat cavernosal smooth muscle (CSM) and the expression of AM system components in this tissue. Functional assays using standard muscle bath procedures were performed in CSM isolated from male Wistar rats. Protein and mRNA levels of pre-pro-AM, calcitonin receptor-like receptor (CRLR), and Subtypes 1, 2 and 3 of the receptor activity-modifying protein (RAMP) family were assessed by Western immunoblotting and quantitative real-time polymerase chain reaction, respectively. Nitrate and 6-keto-prostaglandin F1α (6-keto-PGF1α; a stable product of prostacyclin) levels were determined using commercially available kits. Protein and mRNA of AM, CRLR, and RAMP 1, -2, and -3 were detected in rat CSM. Immunohistochemical assays demonstrated that AM and CRLR were expressed in rat CSM. AM relaxed CSM strips in a concentration-dependent manner. AM22-52, a selective antagonist for AM receptors, reduced the relaxation induced by AM. Conversely, CGRP8-37, a selective antagonist for calcitonin gene-related peptide receptors, did not affect AM-induced relaxation. Preincubation of CSM strips with NG-nitro-L-arginine-methyl-ester (L-NAME, nitric oxide synthase inhibitor), 1H-(1,2,4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, quanylyl cyclase inhibitor), Rp-8-Br-PET-cGMPS (cGMP-dependent protein kinase inhibitor), SC560 [5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-trifluoromethyl pyrazole, selective cyclooxygenase-1 inhibitor], and 4-aminopyridine (voltage-dependent K+ channel blocker) reduced AM-induced relaxation. On the other hand, 7-nitroindazole (selective neuronal nitric oxide synthase inhibitor), wortmannin (phosphatidylinositol 3-kinase inhibitor), H89 (protein kinase A inhibitor), SQ22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, adenylate cyclase inhibitor], glibenclamide (selective blocker of ATP-sensitive K+ channels), and apamin (Ca2+-activated channel blocker) did not affect AM-induced relaxation. AM increased nitrate levels and 6-keto-PGF1α in rat CSM. The major new contribution of this research is that it demonstrated expression of AM and its receptor in rat CSM. Moreover, we provided evidence that AM-induced relaxation in this tissue is mediated by AM receptors by a mechanism that involves the nitric oxide-cGMP pathway, a vasodilator prostanoid, and the opening of voltage-dependent K+ channels.
Descritores: Adrenomedulina/farmacologia
Proteína Semelhante a Receptor de Calcitonina/análise
Músculo Liso/efeitos dos fármacos
Parassimpatolíticos/farmacologia
Pênis/efeitos dos fármacos
Vasodilatadores/farmacologia
-/farmacologia
ABBREVIATIONS AS TOPIC-AMINOPYRIDINE/farmacologia
/análise
ABDOMEN, ACUTE-KETOPROSTAGLANDIN F1 ALPHA/análise
Adrenomedulina/genética
Adrenomedulina/metabolismo
Western Blotting
Proteína Semelhante a Receptor de Calcitonina/antagonistas & inibidores
Proteínas Quinases Dependentes de GMP Cíclico/antagonistas & inibidores
Inibidores de Ciclo-Oxigenase/farmacologia
Relação Dose-Resposta a Droga
Inibidores Enzimáticos/farmacologia
Imuno-Histoquímica
Indazóis/farmacologia
Relaxamento Muscular
Músculo Liso/metabolismo
Óxido Nítrico Sintase/antagonistas & inibidores
Óxido Nítrico/análise
Óxido Nítrico/análogos & derivados
Pênis/metabolismo
Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo
Ratos Wistar
Reação em Cadeia da Polimerase em Tempo Real
RNA Mensageiro/metabolismo
Proteína 1 Modificadora da Atividade de Receptores/genética
Proteína 1 Modificadora da Atividade de Receptores/metabolismo
/metabolismo
RECEPTOR ACTIVITY-MODIFYING PROTEIN ABATTOIRS/metabolismo
/genética
RECEPTOR ACTIVITY-MODIFYING PROTEIN TEMEFOS/genética
/metabolismo
RECEPTOR ACTIVITY-MODIFYING PROTEIN TEMEFOS/metabolismo
Receptores de Peptídeo Relacionado com o Gene de Calcitonina/metabolismo
Limites: Animais
Masculino
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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