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J Appl Physiol 73: 1805-1809, 1992;
8750-7587/92 $5.00
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Journal of Applied Physiology, Vol 73, Issue 5 1805-1809, Copyright © 1992 by American Physiological Society


ARTICLES

Reactive oxygen in skeletal muscle. II. Extracellular release of free radicals

M. B. Reid, T. Shoji, M. R. Moody and M. L. Entman
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.

We have tested the hypothesis that diaphragm muscle fibers release superoxide anion radicals (O2-.) into the extracellular space. Fiber bundles were isolated from rat diaphragm and incubated in Krebs-Ringer solution containing cytochrome c (10(-5) M), a standard assay for O2-.. Bundles were either passive or active, i.e., directly stimulated to contract rhythmically. After 1 h, absorbance of reduced cytochrome c in the incubation medium was measured at 550 nm. Absorbance was greater in medium exposed to passive muscle than in medium without muscle (P < 0.01), indicating O2-. release by passive muscle. Absorbance was greater in medium exposed to active muscle than in that exposed to passive muscle (P < 0.01), an increase inhibited by superoxide dismutase (10(3) U/ml). Active bundles fatigued; bundles developing the lowest final stresses produced the greatest absorbance increases (P < 0.001), suggesting that the magnitude of fatigue was inversely related to O2-. release. We conclude that O2-. is released by diaphragm myocytes into the interstitium and surrounding medium, a process accelerated by fatiguing muscular contractions.


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