Spin trapping of free radical metabolites of carbon tetrachloride in vitro and in vivo: effect of acute ethanol administration.
A single dose of ethanol, when administered 18 hr prior to CCl4, potentiates the hepatotoxicity of the halocarbon. In these studies, spin trapping and electron spin resonance (ESR) spectroscopy methods were utilized to determine whether a single ethanol dose increased the metabolism of CCl4 to free radical intermediates. When hepatic microsomes from ethanol-treated or control rats were incubated with CCl4 and the spin trapping agent alpha-phenyl-N-tert-butylnitrone (PBN), the ESR signal of the trichloromethyl radical adduct of PBN was of similar intensity in both groups. The ethanol dose also failed to induce p-nitrophenol hydroxylase activity. When PBN and CCl4 were administered to rats, liver extracts contained ESR signals resulting primarily from the trichloromethyl radical adduct of PBN, and the signals were of similar intensity in both experimental groups. Higher concentrations of the carbon dioxide anion radical adduct of PBN were detected in plasma samples from ethanol-treated rats. However, when hepatocytes from ethanol-treated and control rats were incubated with PBN and CCl4, ESR signals of the carbon dioxide adduct were of similar intensity. These data suggest that the higher concentrations of the carbon dioxide adduct in the blood of ethanol-treated rats may be explained by early CCl4-induced damage to liver cell membranes, rather than increased rates of formation. The data in this report fail to support the hypothesis that a single dose of ethanol stimulates the hepatic metabolism of CCl4 to the trichloromethyl radical. Alternatively, ethanol may potentiate CCl4 toxicity by affecting some critical metabolic step subsequent to trichloromethyl radical formation.
Reinke
L A
LA
Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Canada.
Towner
R A
RA
Janzen
E G
EG
D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002251:Carbon Tetrachloride / Q000378:metabolism* / Q000493:pharmacokinetics; D002252:Carbon Tetrachloride Poisoning / Q000150:complications; D056486:Chemical and Drug Induced Liver Injury; D003497:Cyclic N-Oxides; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D000431:Ethanol / Q000494:pharmacology*; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism; D007064:L-Iditol 2-Dehydrogenase / Q000097:blood; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D009589:Nitrogen Oxides / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013113:Spin Labels
D003497:Cyclic N-Oxides; D005609:Free Radicals; D009589:Nitrogen Oxides; D013113:Spin Labels; C029217:phenyl-N-tert-butylnitrone; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D002251:Carbon Tetrachloride; D007064:L-Iditol 2-Dehydrogenase; D005978:Glutathione