Induction of free radicals in hepatocytes, mitochondria and microsomes of rats by ochratoxin A and its analogs.
Oxidative damage may be one of the manifestations of cellular damage in the toxicity of ochratoxin A (OA). OA; its three natural analogs, OB, OC and O alpha; and three synthetic analogs, the ethyl amide of OA (OE-OA), O-methylated OA (OM-OA), and the lactone-opened OA (OP-OA) were used to study free radical generation in hepatocytes, mitochondria and microsomes from rats. Electron paramagnetic resonance spectroscopy (EPR) using alpha-(4-pyridyl-1-oxide)-N-tert-butyl nitrone (4-POBN) as a spin trapping agent showed an enhanced free radical generation due to the addition of NADPH to the microsomes. An EPR signal was not observed in the mitochondria and hepatocyte samples when they were treated with a variety of agents. Addition of OM-OA together with NADPH and Fe3+ to the microsomes resulted in a strong EPR signal compared with the other analogs, whereas the signal could be quenched by the addition of catalase. OM-OA does not have a dissociable phenolate group and does not chelate Fe3+. The spin adduct hyperfine splitting constants indicated the presence of alpha-hydroxyethyl radicals resulting from generated hydroxyl radicals, which were trapped by 4-POBN. The results also suggested that the production of hydroxyl radicals by OA does not require a dissociable phenolate group or the prior formation of an OA-Fe complex.
Hoehler
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Department of Animal Science, University of Manitoba, Winnipeg, Canada. hoehler@aninut.uni-kiel.de
Marquardt
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McIntosh
A R
AR
Hatch
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GM
D000818:Animals; D004578:Electron Spin Resonance Spectroscopy; D005260:Female; D005290:Ferric Compounds / Q000494:pharmacology; D005609:Free Radicals; D017665:Hydroxyl Radical / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D009183:Mycotoxins / Q000494:pharmacology*; D009249:NADP / Q000494:pharmacology; D009589:Nitrogen Oxides; D009793:Ochratoxins / Q000494:pharmacology*; D011725:Pyridines; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels; D013482:Superoxide Dismutase / Q000494:pharmacology
D005290:Ferric Compounds; D005609:Free Radicals; D009183:Mycotoxins; D009589:Nitrogen Oxides; D009793:Ochratoxins; D011725:Pyridines; D013113:Spin Labels; C043438:alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone; C025589:ochratoxin A; D017665:Hydroxyl Radical; D009249:NADP; D013482:Superoxide Dismutase