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Intracellular generation of reactive oxygen species during nonhypoxic lung ischemia.

Abstract
Surface fluorometry with 40 microM hydroethidine (HE) as a probe was used to detect oxidant generation in isolated, ventilated rat lungs during lung ischemia. Ethidium fluorescence due to HE oxidation was continuously monitored with 470 nm excitation and 610 nm emission. Fluorescence increased with ischemia in O2-ventilated lungs [0.98 +/- 0.08 arbitrary fluorescence units (AFU)/min vs. 0.58 +/- 0.07 with control perfusion]. HE oxidation during ischemia was prevented by N2 ventilation but was unaltered by preperfusion with superoxide dismutase. Ethidium fluorescence in homogenate prepared from lungs subjected to 1 h of nonhypoxic ischemia was increased (16.8 +/- 1.5 vs. 9.8 +/- 0.4 AFU/mg protein in control) but was unchanged in lungs that had been N2 ventilated. Microfluorographs of HE perfused and fixed lung sections demonstrated marked generalized increases in ethidium fluorescence with ischemia compared with control perfusion. Ischemia resulted in significant increases in tissue thiobarbituric acid reactive substance (176 +/- 13 vs. 44 +/- 3 pmol/mg protein for control) and in lung conjugated dienes (0.90 +/- 0.07 vs. 0.48 +/- 0.06 U/mg protein for control), indicating peroxidation of lung lipids. These results indicate that lung ischemia leads to intracellular oxidant generation that can be continuously monitored by surface fluorometry.
AuthorsA B Al-Mehdi, H Shuman, A B Fisher
JournalThe American journal of physiology (Am J Physiol) Vol. 272 Issue 2 Pt 1 Pg. L294-300 (Feb 1997) ISSN: 0002-9513 [Print] United States
PMID9124381 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Lipid Peroxides
  • Phenanthridines
  • Reactive Oxygen Species
  • hydroethidine
Topics
  • Animals
  • Fluorometry
  • In Vitro Techniques
  • Intracellular Membranes (metabolism)
  • Ischemia (metabolism)
  • Lipid Peroxides (metabolism)
  • Male
  • Phenanthridines (pharmacology)
  • Pulmonary Circulation
  • Rats
  • Rats, Sprague-Dawley
  • Reactive Oxygen Species (metabolism)

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