Mechanisms underlying the vasorelaxant action of the pimarane ent-8(14),15-pimaradien-3beta-ol in the isolated rat aorta.

摘要

Pimarane-type diterpenes were described to exert antispasmodic and relaxant activities. Based on this observation we hypothesized that the diterpene ent-8(14),15-pimaradien-3beta-ol (PA-3beta-ol) induced vascular relaxation. With this purpose, the present work investigates the mechanisms involved in the vasorelaxant effect of the pimarane-type diterpene PA-3beta-ol. Vascular reactivity experiments, using standard muscle bath procedures, were performed in isolated aortic rings from male Wistar rats. Cytosolic calcium concentration ([Ca(2+)]c) was measured by confocal microscopy using the fluorescent probe Fluo-3AM. PA-3beta-ol (10, 50 and 100 micromol/l) inhibited phenylephrine and KCl-induced contraction in either endothelium-intact or denuded rat aortic rings. PA-3beta-ol also reduced CaCl(2)-induced contraction in Ca(2+)-free solution containing KCl (30 mmol/l) or phenylephrine (0.1 micromol/l). PA-3beta-ol (1-300 micromol/l) concentration dependently relaxed phenylephrine-pre-contracted rings with intact or denuded endothelium. The diterpene also relaxed KCl-pre-contracted rings with intact or denuded endothelium. Moreover, Ca(2+) mobilization study showed that PA-3beta-ol (100 micromol/l) and verapamil (1 micromol/l) inhibited the increase in Ca(2+)-concentration in smooth muscle and endothelial cells induced by phenylephrine (10 micromol/l) or KCl (60 mmol/l). Pre-incubation of intact or denuded aortic rings with N(G)-nitro-l-arginine methyl ester (L-NAME, 100 micromol/l) and 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 1 micromol/l) produced a rightward displacement of the PA-3beta-ol concentration-response curves. On the other hand, 7-nitroindazole (100 micromol/l), 1400 W (1 micromol/l), indomethacin (10 micromol/l) and tetraethylammonium (1 mmol/l) did not affect PA-3beta-ol-induced relaxation. Collectively, our results provide evidence that the effects elicited by PA-3beta-ol involve extracellular Ca(2+) influx blockade. Its effects are also partly mediated by the activation of NO-cGMP pathway.