Action of Reactive Oxygen Species and Their Antagonists on Twitch Tension of the Rat Phrenic Nerve‐Diaphragm

Abstract

Reactive oxygen species have been implicated in normal and pathological processes of many tissues, including skeletal muscle. I extended previous studies by examining the effect of these intermediates and eight of their antagonists (superoxide dismutase, catalase, deferoxamine, [Cu(II)]2(3,5-diisopropylsalicylate)4, 1,2-dimethyl-3-hydroxy-pyridone, 1,3-dimethyl-2-thiourea, N-(2-mercaptopropionyl)-glycine, vitamin E) on indirectly stimulated twitch tension of an in vitro neuroskeletomuscular preparation, the phrenic nerve-diaphragm of the rat. In the absence of exogenous reactive oxygen species, none of the antagonists potentiated twitch tension, and all but one (N-[2-mercaptopropionyl]-glycine) of the membrane-permeant antagonists attenuated twitch tension. The reactive oxygen intermediate-generating system of purine plus xanthine oxidase reduced indirectly stimulated twitch tension by 36% while having no effect on directly stimulated twitch tension. Catalase (but not superoxide dismutase or deferoxamine) eliminated the reduction in twitch tension, indicating that hydrogen peroxide played a role in the reduction. The membrane-permeant antagonists [Cu(II)]2(3,5-diisopropylsalicylate)4 and 1,2-dimethyl-3-hydroxy-pyridone also eliminated the reduction in twitch tension caused by reactive oxygen species, suggesting that hydrogen peroxide could have acted intracellularly through an iron-catalyzed Haber-Weiss reaction to produce hydroxyl radical, which in turn reacted with intracellular components, thereby reducing twitch tension.