C2-ceramide attenuates phenylephrine-induced vasoconstriction and elevation in [Ca2+]i in rat aortic smooth muscle.

In the present study, we examined the effects of cell-permeable C2-ceramide on contraction of aortic smooth muscle and intracellular free Ca2+ ([Ca2+]i). C2-ceramide (10(-7) to 10(-4) M) alone did not elicit any significant changes in either basal tension or resting levels of [Ca2+]i in rat aortic smooth muscle. However, C2-ceramide (10(-7) to 10(-4) M) attenuated phenylephrine-induced contractions in isolated rat aortic rings in a concentration-related manner, and inhibited elevations in [Ca2+]i in cultured rat aortic smooth muscle cells induced by phenylephrine. C2-ceramide-induced relaxation was found to be only slightly endothelium-dependent. However, nitric oxide inhibitors (L-NNA, L-NMMA), an inhibitor of prostanoid synthesis (indomethacin), an inhibitor of opiate actions, and several inhibitors of the pharmacologic actions of various vasoactive amines all failed to interfere with the vasorelaxant responses of C2-ceramide. Three different inhibitors of protein kinase C, when used in a wide concentration range, also failed to interfere with the ceramide-induced relaxations. Our results suggest that the sphingomyelin-signaling pathway may play an important regulatory role in arterial wall tone.