Mechanisms underlying the hydrogen peroxide-induced, endothelium-independent relaxation of the norepinephrine-contraction in guinea-pig aorta.

摘要

The mechanisms underlying the hydrogen peroxide-induced relaxation of the norepinephrine-contraction were studied by measuring isometric force, myosin light chain (MLC(20)) phosphorylation and cyclic GMP in endothelium-denuded muscle from the guinea-pig aorta. Norepinephrine (5.2+/-1.3 microM) produced a phasic, followed by a tonic contraction. Hydrogen peroxide (10 and 100 microM), glyceryl trinitrate (30 and 300 nM) and 8-bromo cyclic GMP (30 and 100 microM) did not change the basal tone, but reduced the norepinephrine-induced contraction. Phosphorylation of MLC(20) (percentage of phosphorylated to total MLC(20)) was increased 1 min (5.9+/-1.0% vs. 35.9+/-4.9%) and, to a lesser extent, 20 min (3.7+/-1.7% vs. 13.9+/-1.6%) after the addition of norepinephrine. Hydrogen peroxide (100 microM) did not modify basal MLC(20) phosphorylation, but reduced the increase in MLC(20) phosphorylation induced by 1-min exposure to norepinephrine (20.9+/-4.1%). Its effect was abolished by catalase. When the tissue was incubated for 20 min with norepinephrine in the presence of hydrogen peroxide, norepinephrine-induced MLC(20) phosphorylation was not changed (13.6+/-1.5%), as compared to that in the absence of hydrogen peroxide. Hydrogen peroxide relaxed norepinephrine-stimulated aortas in a concentration-dependent fashion with EC(50) values of 5.9+/-0.2 microM. The relaxation was inhibited by soluble guanylate cyclase inhibitors and increased by an inhibitor of cyclic GMP-selective phosphodiesterase. In aorta precontracted with norepinephrine, hydrogen peroxide (100 microM) relaxed the tissue by 89+/-11% and almost doubled tissue concentrations of cyclic GMP, whereas sodium nitroprusside (1 microM) relaxed the tissue by 100% and increased cyclic GMP concentrations 30-fold. It is suggested that the inhibitory effects of hydrogen peroxide on the norepinephrine-induced phasic and sustained contractions are explained by a decrease in MLC(20) phosphorylation and by an alteration in MLC(20) phosphorylation-independent mechanisms, respectively. The effects of hydrogen peroxide were in part mediated by cyclic GMP.