Decreased contraction to phenylephrine by ouabain in 2K-1C rat aorta is modulated by the endothelium.

The effects of ouabain were studied on the contraction stimulated with phenylephrine or KCl in intact endothelium and denuded aortic rings isolated from normotensive (2K) and renal hypertensive 2 kidney-1clip (2K-1C) rats. Ouabain did not change the basal tone of aortic rings. Ouabain (1 nmol/l) had no effect on the contraction to phenylephrine in all the artery groups studied. Ouabain (10 nmol/l) decreased the E(max) to phenylephrine in intact endothelium arteries from 2K-1C. By contrast, ouabain (10 nmol/l) had no effect on the contraction to KCl. Ouabain induced membrane depolarization measured by confocal image with Di-4-ANEPPS dye, that was greater in 2K than in 2K-1C rat aorta smooth muscle cells. In conclusion, ouabain (10 nmol/l) decreased the contractile responses to phenylephrine only in 2K-1C rat aortic rings with intact endothelium. Interestingly, 10 nmol/l ouabain depolarizes the smooth muscle cells but this depolarization level is not enough to alter the phenylephrine or KCl-induced contractions. Our results indicate that the endothelium modulates the vascular action of ouabain.

Clonidine induces rat aorta relaxation by nitric oxide-dependent and -independent mechanisms.

The alpha1- and alpha2-adrenoceptors coexist in vascular smooth muscle cells producing vascular contraction and relaxation. This study was designed to investigate which is the mechanism activated by clonidine in the rat aorta, and the endothelial factors possibly involved in the relaxation induced by clonidine. The alpha2-adrenoceptors agonist clonidine relaxed rat aortas pre-contracted with phenylephrine, with or without endothelium. In non-contracted denuded arteries, clonidine produced contractions instead of relaxation. In intact endothelium aortic rings, clonidine induced greater relaxation than in denuded aortic rings. In aortas with intact endothelium, the NO-synthase inhibitor L-NAME (10 micromol/L) and the NO-scavenger hemoglobin (10 micromol/L) reduced the relaxation to clonidine. On the other hand, indomethacin (10 micromol/L) failed to alter the relaxation induced by clonidine. These results suggest the participation of NO, but not prostacyclin in clonidine-induced relaxation. In aortic rings pre-contracted with KCl (60 mmol/L) the relaxation induced by clonidine was abolished; however, the K+ channel blockers glibenclamide (K(ATP)), tetraethylamonium (K(Ca)), and the combination of apamin and charybdotoxin (K(Ca)) did not change the relaxation induced by clonidine. The relaxation induced by clonidine on PGF2alpha-contracted arteries was not affected by prazosin. However, in the absence of prazosin, clonidine had an additional contractile effect in PGF2alpha-contracted arteries. In conclusion, our results show that in rat aorta clonidine can activate alpha2-adrenoceptors in the smooth muscle cells and alpha2-adrenoceptors in the endothelial cells that activates NO production, but not prostacyclin and/or EDHF. In the absence of phenylephrine and prazosin, clonidine can also activate alpha1-adrenoceptors and rat aorta contraction.