Nitric oxide relaxes human myometrium by a cGMP-independent mechanism

The role of intracellular guanosine 3',5'-cyclic monophosphate concentratio n ([cGMP](i)) in nitric oxide (NO)-mediated relaxations in the uterus has b ecome controversial. We found the NO donor S-nitroso-L-cysteine (CysNO) to potently (IC50 = 30 nM) inhibit spontaneous contractions in the nonpregnant human myometrium. CysNO treatment increased [cGMP](i) significantly (P < 0 .001), and this increase was blocked by the guanylyl cyclase inhibitors met hylene blue (10 mu M) or LY-83583 (1 mu M); however, pretreatment with thes e guanylyl cyclase inhibitors failed to block CysNO-mediated relaxations. I ntracellular cAMP concentrations were not altered by treatment of tissues w ith 10 mu M CysNO. Incubation with the cGMP analogs 8-bromo-cGMP or beta-ph enyl-1,N-2-etheno-cGMP did not significantly affect spontaneous contractili ty. Pretreatment of tissues with charybdotoxin [a calcium-dependent potassi um channel (BK) blocker] completely reversed CysNO-induced relaxations. We conclude that NO is a potent inhibitor of spontaneous contractile activity in the nonpregnant human uterus and that, although guanylyl cyclase and BK activities are increased by NO, increases in [cGMP](i) are not required for NO-induced relaxations in this tissue.