The tension of isolated ring preparation of aorta from nonpregnant and pregnant rats was measured isometrically to study the effect of pregnancy on endothelium-derived relaxing factor activity. Contraction in response to norepinephrine and potassium chloride was greater in aortae from nonpregnant rats than in those from pregnant rats. The endothelium-dependent relaxation that was caused by acetylcholine (10(-10)-3 x 10(-9) M) in aortae precontracted with norepinephrine was significantly enhanced in aortae from pregnant rats compared with the relaxation in those from nonpregnant rats. NG-nitro-L-arginine methyl ester (L-NAME) inhibited the endothelium-dependent relaxation in both aorta from pregnant and nonpregnant rats. L-Arginine reversed the inhibition of L-NAME. Those results suggest that the enhanced endothelium-derived relaxing factor activity in rats aortae is associated with pregnancy.