Adenosine relaxation in isolated rat aortic rings and possible roles of smooth muscle Kv channels, KATP channels and A2a receptors

Background: An area of ongoing controversy is the role adenosine to regulate vascular tone in conduit vessels that regulate compliance, and the role of nitric oxide (NO), potassium channels and receptor subtypes involved. The aim of our study was to investigate adenosine relaxation in rat thoracic aortic rings, and the effect of inhibitors of NO, prostanoids, K-v, K-ATP channels, and A(2a) and A(2b) receptors. Methods: Aortic rings were freshly harvested from adult male Sprague Dawley rats and equilibrated in an organ bath containing oxygenated, modified Krebs-Henseleit solution, 11 mM glucose, pH 7.4, 37 degrees C. Isolated rings were pre-contracted sub-maximally with 0.3 mu M norepinephrine (NE), and the effect of increasing concentrations of adenosine (1 to 1000 mu M) were examined. The drugs L-NAME, indomethacin, 4-aminopyridine (4-AP), glibenclamide, 5-hydroxydecanoate, ouabain, 8-(3-chlorostyryl) caffeine and PSB-0788 were examined in intact and denuded rings. Rings were tested for viability after each experiment. Results: Adenosine induced a dose-dependent, triphasic relaxation response, and the mechanical removal of the endothelium significantly deceased adenosine relaxation above 10 mu M. Interestingly, endothelial removal significantly decreased the responsiveness (defined as % relaxation per mu M adenosine) by two-thirds between 10 and 100 mu M, but not in the lower (1-10 mu M) or higher (>100 mu M) ranges. In intact rings, L-NAME significantly reduced relaxation, but not indomethacin. Antagonists of voltage-dependent K-v (4-AP), sarcolemma K-ATP (glibenclamide) and mitochondrial K-ATP channels (5-HD) led to significant reductions in relaxation in both intact and denuded rings, with ouabain having little or no effect. Adenosine-induced relaxation appeared to involve the A(2a) receptor, but not the A(2b) subtype. Conclusions: It was concluded that adenosine relaxation in NE-precontracted rat aortic rings was triphasic and endothelium-dependent above 10 mu M, and relaxation involved endothelial nitric oxide (not prostanoids) and a complex interplay between smooth muscle A(2a) subtype and voltage-dependent K-v, SarcK(ATP) and MitoK(ATP) channels. The possible in vivo significance of the regulation of arterial compliance to left ventricular function coupling is discussed.