Evidence for the presence of a microsomal NADH-dependent enzyme system that can bioactivate aromatic amines in the liver of rats and mice.
Experimental evidence is presented for the presence in the liver of rats and mice of an Aroclor 1254-inducible, NADH-dependent enzyme system that can catalyse the bioactivation of aromatic and heterocyclic amines to genotoxic metabolites. It differs from the established microsomal cytochrome P450 and flavin monooxygenase systems in its response to treatment with cytochrome P450 inducing agents, optimum protein concentration and in vitro modulation by DMSO. The mutagenic metabolites generated by the NADH-supported system appear to be similar to those generated by the NADPH-mediated systems. Mutagenicity of the aminocompounds in the presence of either cosubstrate was less pronounced in an O-acetyltransferase-deficient bacterial strain, implying the presence of hydroxylamines. Moreover, glutathione potentiated the mutagenic response of both the NADH- and NADPH-generated metabolites. Cytochrome c suppressed markedly the NADPH-dependent mutagenicity of aromatic amines but had no such effect in the presence of NADH. Similarly, antibodies to cytochrome P450 reductase markedly inhibited the NADPH-, but not the NADH-dependent bioactivation of the aromatic amine 2-aminoanthracene. The cytochrome P450 suicide inhibitor, 1-aminobenzotriazole, decreased the mutagenicity of both, the NADH- and NADPH-mediated bioactivation of the aminocompounds. The above findings raise the possibility that a cytochrome P450-like protein, that can receive electrons from NADH, possibly through cytochrome b5 reductase, is present in the hepatic microsomes of rats and mice, and is capable of catalysing the bioactivation of aromatic amines through N-hydroxylation. Such a hypothesis is supported by the findings that NADH could support the O-dealkylation of 7-methoxy- and 7-ethoxy-resorufin, in the absence of NADPH. Finally the NADH-dependent bioactivation of aromatic amines was induced markedly by Aroclor 1254 and benzo(a)pyrene in Ah responsive, but not Ah nonresponsive, mice indicating that it is associated with the Ah locus.
Molecular Toxicology Group, School of Biological Sciences, University of Surrey, Guildford, UK.
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Marczylo
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Ioannides
D000818:Animals; D000873:Anthracenes / Q000493:pharmacokinetics* / Q000633:toxicity; D001711:Biotransformation; D002273:Carcinogens / Q000493:pharmacokinetics*; D003577:Cytochrome P-450 Enzyme System / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008862:Microsomes, Liver / Q000378:metabolism*; D009153:Mutagens / Q000493:pharmacokinetics; D009243:NAD / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
D000873:Anthracenes; D002273:Carcinogens; D009153:Mutagens; D009243:NAD; C006593:2-anthramine; D003577:Cytochrome P-450 Enzyme System