To determine whether amrinone and its induced increase of cyclic AMP releases nitric oxide and enhances endothelium-dependent vascular relaxation, we studied nitric oxide production and vascular relaxation of rat thoracic aorta on treatment with amrinone and forskolin, an activator of adenylate cyclase. When 20 microM amrinone was applied to ring segments of aorta previously contracted with phenylephrine, relaxation was greater in segments with endothelium than in those without (% relaxation 94 +/- 4 vs 37 +/- 7%, P < 0.01). However, a higher concentration of amrinone (> 50 microM) induced the same degree of relaxation in ring segments with or without endothelium, probably due to its direct effect on vascular smooth muscle. The maximal relaxant concentration (100 microM) of amrinone induced an increase (2.5 fold) in cyclic GMP in ring segments. Forskolin also induced concentration-dependent relaxation, but removal of the endothelium attenuated the relaxation induced by forskolin about seven-fold. NG-nitro L-arginine reversed the relaxation induced by amrinone or forskolin in ring segments with, but not without, endothelium. Nitric oxide production in ring segments of aorta, following application of amrinone or forskolin, was detected by nitric oxide-selective electrode and electron paramagnetic resonance spin trapping methods. Pre-treatment with NG-nitro L-arginine or removal of the endothelium suppressed nitric oxide production by amrinone or forskolin. These data showed that amrinone enhances the release of nitric oxide from rat aortic endothelial cells, and induces endothelium-derived relaxing factor/nitric oxide-mediated vasodilation.