Ferrer Mercedes, Sánchez Margarita, Martín Maria del Carmen, Márquez-Rodas Iván, Alonso Maria Jesús, Salaices Mercedes, Balfagón Gloria
Departamento de Fisiología, Facultad de Medicina, Universidad Autónoma, C/Arzobispo Morcillo, 4, Madrid 28029, Spain.
Eur J Pharmacol. 2004 Mar 8;487(1-3):167-73. doi: 10.1016/j.ejphar.2004.01.030.
The aim of this study was to analyse the possible influence of cyclic AMP-protein kinase A (cAMP-PKA) activation on neuronal nitric oxide (NO) release induced by electrical field stimulation in mesenteric arteries from Wistar Kyoto (WKY) rats. Western blot experiments demonstrated the expression of neuronal NO synthase (nNOS) in mesenteric artery from WKY rats; however, electrical field stimulation alone did not induce detectable NO release. Preincubation with forskolin allowed NO release induced by electrical field stimulation, which was abolished by: the neuronal toxine tetrodotoxin, the nNOS inhibitors 7-nitroindazole or N(omega)-propil-l-arginine (NPLA), and the PKA inhibitors N-(2-(p-Bromocinnamylamino) ethyl 5-isoquinolinesulfonamide hydrochloride (H-89) or (9R,10S,12S)-2,3,9,10,11, 12-Hexahydro-10-9-methyl-1-oxo-9,12-epoxy-1H-diindolo(1,2,3-fg:3,2,1k)pyrrolo(3,4-l)(1,6) benzodiazocine-10-carboxylic acid hexyl ester (KT-5720). Preincubation with prostacyclin also allowed the NO release induced by electrical field stimulation which was significantly decreased by: the neuronal toxine tetrodotoxin, the nNOS inhibitors 7-nitroindazole or NPLA, and the PKA inhibitors H-89 or KT-5720. The NOS inhibitor N(omega)-nitro-l-arginine methyl ester (l-NAME) did not modify the vasoconstrictor response induced by electrical field stimulation. However, in the presence of forskolin or prostacyclin, l-NAME increased the vasoconstrictor response to electrical field stimulation. These results indicate that forskolin and prostacyclin allow neuronal NO release induced by electrical field stimulation through a mechanism involving cAMP-PKA activation in rat mesenteric arteries.
本研究的目的是分析环磷酸腺苷 - 蛋白激酶A（cAMP - PKA）激活对Wistar Kyoto（WKY）大鼠肠系膜动脉电场刺激诱导的神经元型一氧化氮（NO）释放的可能影响。蛋白质免疫印迹实验证明了WKY大鼠肠系膜动脉中神经元型一氧化氮合酶（nNOS）的表达；然而，单独的电场刺激并未诱导出可检测到的NO释放。用福斯可林预孵育可使电场刺激诱导的NO释放，而这种释放可被以下物质消除：神经毒素河豚毒素、nNOS抑制剂7 - 硝基吲唑或N（ω） - 丙基 - L - 精氨酸（NPLA），以及PKA抑制剂N - （2 - （对 - 溴肉桂氨基）乙基） - 5 - 异喹啉磺酰胺盐酸盐（H - 89）或（9R，10S，12S） - 2，3，9，10，11，12 - 六氢 - 10 - 9 - 甲基 - 1 - 氧代 - 9，12 - 环氧 - 1H - 二吲哚并（1，2，3 - fg：3，2，1k）吡咯并（3，4 - l）（1，6）苯并二氮杂䓬 - 10 - 羧酸己酯（KT - 5720）。用前列环素预孵育也可使电场刺激诱导的NO释放，而这种释放可被以下物质显著降低：神经毒素河豚毒素、nNOS抑制剂7 - 硝基吲唑或NPLA，以及PKA抑制剂H - 89或KT - 5720。一氧化氮合酶抑制剂N（ω） - 硝基 - L - 精氨酸甲酯（L - NAME）并未改变电场刺激诱导的血管收缩反应。然而，在存在福斯可林或前列环素的情况下，L - NAME增强了对电场刺激的血管收缩反应。这些结果表明，福斯可林和前列环素通过一种涉及大鼠肠系膜动脉中cAMP - PKA激活的机制，使电场刺激诱导神经元型NO释放。
