Modulation of aflatoxin B1 biotransformation in rabbit pulmonary and hepatic microsomes.
Aflatoxin B1 (AFB1) is a carcinogenic mycotoxin that requires activation to the corresponding 8,9-epoxide for activity. In addition to being present in foodstuffs, AFB1 can contaminate respirable grain dusts and thus the respiratory system is a potential target for carcinogenesis. In the present study, we have investigated the role of polycyclic aromatic hydrocarbon-inducible forms of cytochrome P-450 in the pulmonary and hepatic microsomal activation ([3H]AFB1-DNA binding) and detoxification ([3H]AFM1 and [3H]AFQ1 formation) of [3H]AFB1. In rabbit lung microsomes, the apparent Vmax for [3H]AFM1 formation was increased significantly when values were expressed per mg microsomal protein or per nmol P-450 present. In liver microsomes, the apparent Vmax for DNA binding and [3H]AFM1 formation were increased by beta-naphthoflavone (BNF) treatment (to 2.3 and 3.3 times control, respectively) when expressed per mg protein, but when expressed per nmol P-450, only AFM1 formation was significantly increased. The apparent Km values for both these reactions were unaffected. The apparent Vmax for [3H]AFQ1 formation was not affected by BNF treatment, but the apparent Km was increased to 4.5 times control. Boiling of microsomes or omitting the NADPH-generating system decreased DNA binding, AFM1 formation and AFQ1 formation by 89-97%, while addition of 1.0 mM SKF-525A inhibited these reactions by 46-57%. Addition of 1.0 mM alpha-naphthoflavone (ANF) had no effect on the biotransformation of [3H]AFB1 in lung microsomes of control rabbits, but significantly decreased AFM1 formation (by 31%) in lung microsomes from BNF-treated animals (other reactions were unaffected). In liver microsomes from BNF treated rabbits, 1.0 mM ANF inhibited DNA binding of [3H]AFB1 by 68%, while there was no effect in control microsomes. ANF significantly inhibited AFM1 formation in liver microsomes from both control and BNF-treated animals (by 87-97% and 67-78% at 1.0 mM and 2.0 microM, respectively), but had no effect on AFQ1 formation in liver microsomes from animals in either treatment group. These results indicate an important role for the 1A subclass of P-450 isozymes in the biotransformation of AFB1 to AFM1 in rabbit lung and liver, and a minor role in AFB1 activation in liver.
Daniels
J M
JM
Department of Pharmacology, Queen's University, Kingston, Ontario, Canada.
Massey
T E
TE
D016604:Aflatoxin B1 / Q000378:metabolism / Q000493:pharmacokinetics*; D016607:Aflatoxin M1 / Q000096:biosynthesis; D000348:Aflatoxins / Q000096:biosynthesis / Q000378:metabolism; D000818:Animals; D001571:Benzoflavones / Q000494:pharmacology; D001711:Biotransformation; D019363:Cytochrome P-450 CYP1A1; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004790:Enzyme Induction; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000187:drug effects; D007700:Kinetics; D008168:Lung / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000187:drug effects / Q000378:metabolism; D011335:Proadifen / Q000494:pharmacology; D011817:Rabbits; D019324:beta-Naphthoflavone
D000348:Aflatoxins; D001571:Benzoflavones; D065607:Cytochrome P-450 Enzyme Inhibitors; D007527:Isoenzymes; C012170:aflatoxicol; C033759:aflatoxin Q1; D009249:NADP; C011512:alpha-naphthoflavone; D019324:beta-Naphthoflavone; D016607:Aflatoxin M1; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1; D011335:Proadifen; D010088:Oxidoreductases; D019363:Cytochrome P-450 CYP1A1