Binding of nitropyrenes and benzo[a]pyrene to mouse lung deoxyribonucleic acid after pretreatment with inducing agents

Abstract In assessing the biological effects of exposure to a complex chemical mixture, it is important to determine how the behavior of one compound may be influenced by the presence of other compounds in the mixture. In this study the effect of pre-exposure to an organic extract of diesel exhaust or to selected compounds in diesel exhaust on the binding of diesel exhaust compounds to DNA was determined. The amount of radiolabel covalently bound to mouse lung DNA following intratracheal administration of radiolabeled benzo[ a ]pyrene (BaP), 1-nitropyrene, 1,3,6-trinitropyrene, or a mixture of dinitropyrene was determined following pretreatment with benzo[ a ]pyrene, 1-nitropyrene, and diesel exhaust extract. Male CD-1 mice, 15–18 weeks of age, received 10 mg/kg radiolabeled putative inducing agents by intratracheal instillation and, after 24 hr, 0.03 to 1.2 mg/kg radiolabeled putative DNA binding agents. Lung DNA was extracted, and covalent binding was quantitated by liquid scintillation spectroscopy. 1-Nitropyrene was a potent lung DNA binding agent in the absence of inducing agents [Covalent Binding Index (CBI) = 970] and was extremely potent after benzo[a]pyrene pretreatment (CBI = 21,540, comparable to the CBI for aflatoxin B 1 ). Similar results were obtained for DNA binding of dinitropyrene and trinitropyrene with and without BaP pretreatment. DNA binding of BaP was lower (CBI = 40) and less inducible (BaP-pretreatment CBI = 230). Pretreatment with diesel extract caused an elevation in the binding of benzo[ a ]pyrene but little or no elevation in the binding of the nitropyrenes. Pretreatment with 1-nitropyrene did not increase significantly DNA binding of any of the agents tested. These results indicate that nitropyrenes bind readily to lung DNA and this binding may be increased in the presence of respirable mixtures, especially those containing inducing agents such as BaP.