This study investigates the participation of the endothelial factors in the alpha-adrenoceptor contractile responses in mesenteric resistance arteries from 15 days ouabain-treated (25 microg/kg/day) and untreated rats. Ouabain treatment increased blood pressure and heart rate without changing the contractile response to phenylephrine (3 nM-30 microM). Endothelium removal or N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM), increased the responses to phenylephrine. The endothelial modulation was similar in both rat groups, but the L-NAME effects were bigger in arteries from ouabain-treated rats. However, the endothelial NOS expression and the relaxation to acetylcholine (0.1 nM-10 microM) remained unaltered after ouabain treatment. The coincubation with L-NAME and indomethacin (100 microM) leftward shifted the concentration-response curves to phenylephrine in arteries from untreated rats similarly to the displacement after incubation only with L-NAME. However, in mesenteric arteries from treated rats, the co-incubation with indomethacin and L-NAME did not alter the response to phenylephrine. The addition of the inhibitor of calcium activated potassium channels tetraethylammonium (2 mM) further leftward shifted the phenylephrine curves only in arteries from untreated rats. Cyclooxygenase-2 (COX-2) expression was greater in vessels from ouabain-treated rats. In conclusion, the chronic ouabain treatment for 15 days modified the participation of endothelial factors in response to phenylephrine in mesenteric resistance arteries, by increasing the release of NO and prostanoids and impairment the endothelium-derived hyperpolarizing factor (EDHF) release. This was accompanied by an increased COX-2 expression. Although this balance avoids changes in the phenylephrine concentration-response curves, these vascular changes might contribute to maintain the ouabain-induced hypertension.