Metabolism of acetaldehyde to methyl and acetyl radicals: in vitro and in vivo electron paramagnetic resonance spin-trapping studies.
Acetaldehyde oxidation by enzymes and cellular fractions has been previously shown to produce radicals that have been characterized as superoxide anion, hydroxyl, and acetyl radicals. Here, we report that acetaldehyde metabolism by xanthine oxidase, submitochondrial particles and whole rats produces both the acetyl and the methyl radical, although only the latter was unambiguously identified in vivo. Electron paramagnetic resonance (EPR) characterization of both radicals was possible by the use of two spin traps, 5,5-dimethyl 1-pyrroline N-oxide (DMPO) and alpha-(4-pyridyl 1-oxide)-N-t-butylnitrone (POBN), and of acetaldehyde labeled with (13)C. The POBN-acetyl radical adduct proved to be unstable, but POBN was employed to monitor acetaldehyde metabolism by Sprague-Dawley rats because previous studies have shown its usefulness for in vivo spin trapping. EPR analysis of the bile collected from treated and control rats showed the presence of the POBN-methyl and of an unidentified, biomolecule-derived, POBN adduct. Because decarbonylation of the acetyl radical is one of the routes for methyl radical formation from acetaldehyde, detection of the latter in bile provides strong evidence for the production of both radicals in vivo. The results may be relevant to understanding the toxic effects of acetaldehyde itself and of its more relevant biological precursor, ethanol.
Nakao
L S
LS
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
Kadiiska
M B
MB
Mason
R P
RP
Grijalba
M T
MT
Augusto
O
O
D000079:Acetaldehyde / Q000378:metabolism*; D000818:Animals; D001646:Bile / Q000378:metabolism; D002417:Cattle; D003497:Cyclic N-Oxides; D004492:Edetic Acid / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D005290:Ferric Compounds / Q000378:metabolism; D005609:Free Radicals / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008697:Methane / Q000031:analogs & derivatives / Q000378:metabolism; D008929:Mitochondria, Heart / Q000378:metabolism; D009589:Nitrogen Oxides; D010084:Oxidation-Reduction; D011725:Pyridines; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels; D013367:Submitochondrial Particles / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism
D003497:Cyclic N-Oxides; D005290:Ferric Compounds; D005609:Free Radicals; D009589:Nitrogen Oxides; D011725:Pyridines; D013113:Spin Labels; C043438:alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone; C051224:methyl radical; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D004492:Edetic Acid; D014969:Xanthine Oxidase; D000079:Acetaldehyde; C019179:Fe(III)-EDTA; D008697:Methane