Activated polymorphonuclear leukocytes increase low-level chemiluminescence of isolated perfused rat lungs.
Low-level chemiluminescence was measured in isolated perfused rat lungs subjected to different types of oxidative stress: perfusion with tert-butyl hydroperoxide (t-BOOH) or stimulation of polymorphonuclear cells (PMN). The time required for t-BOOH-dependent lung chemiluminescence to return to background levels was proportional to the concentration of t-BOOH. From the half times of the decay at different t-BOOH concentrations, we estimated that the lungs metabolize organic peroxides at a rate of 0.045 mM/min. Use of a high dose of t-BOOH (3 mM)or pretreatment of lungs with 1,3-bis(2-chloroethyl)-nitrosourea (100 micrograms/ml) to inhibit glutathione reductase produced chemiluminescence that was much greater and did not decay. Stimulation of 5 x 10(7) PMN with 1 micrograms of phorbol myristate acetate resulted in significant increases in chemiluminescence that occurred in the absence of a significant lung weight gain or measurable lipid peroxidation. Perfusion of isolated lungs with superoxide dismutase (100 U/ml) completely inhibited the chemiluminescence response to PMN activation, whereas treatment with 100 microM U-74389F, a lipid-soluble antioxidant, also significantly decreased PMN-dependent chemiluminescence. Neither catalase (2,000 U/ml) nor 100 microM U-78518F, a water-soluble antioxidant, decreased chemiluminescence after PMN activation. These results indicate that low-level chemiluminescence is a sensitive indicator of oxidative stress in the isolated perfused rat lung and provides a tool for devising and characterizing the effectiveness of antioxidant interventions.
Barnard
M L
ML
Department of Biomedical Sciences, University of South Alabama, Mobile 36688.
Gurdian
S
S
Turrens
J F
JF
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002330:Carmustine / Q000494:pharmacology; D003911:Dextrans / Q000494:pharmacology; D016166:Free Radical Scavengers; D005979:Glutathione Peroxidase / Q000494:pharmacology; D066298:In Vitro Techniques; D015227:Lipid Peroxidation / Q000187:drug effects; D008163:Luminescent Measurements; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D010545:Peroxides / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013312:Stress, Physiological / Q000503:physiopathology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism; D020122:tert-Butylhydroperoxide
D000975:Antioxidants; D003911:Dextrans; D016166:Free Radical Scavengers; D016877:Oxidants; D010545:Peroxides; D017392:Thiobarbituric Acid Reactive Substances; D020122:tert-Butylhydroperoxide; D005979:Glutathione Peroxidase; D013755:Tetradecanoylphorbol Acetate; D002330:Carmustine