The impact of glucuronidation on the bioactivation and DNA adduction of the cooked-food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in vivo.
UDP-glucuronosyltransferases (UGTs) catalyze the glucuronidation of many different chemicals. Glucuronidation is especially important for detoxifying reactive intermediates from metabolic reactions, which otherwise can be biotransformed into highly reactive cytotoxic or carcinogenic species. Detoxification of certain food-borne-carcinogenic heterocyclic amines (HAs) is highly dependent on UGT1A-mediated glucuronidation. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), the most mass abundant carcinogenic HA found in well-done cooked meat, is extensively glucuronidated by UGT1A proteins. In humans, CYP1A2 catalyzed N-hydroxylation and subsequent UGT1A-mediated glucuronidation is a dominant pathway in the metabolism of PhIP. Therefore, changes in glucuronidation rates could significantly alter PhIP metabolism. To determine the importance of UGT1A-mediated glucuronidation in the biotransformation of PhIP, hepatic UGT1A deficient Gunn and UGT1A proficient Wistar rats were exposed to a 100 microg/kg oral dose of [(14)C]PhIP. Urine was collected over 24 h and the PhIP urinary metabolite profiles were compared between the two strains. After the 24 h exposure, livers and colons were removed and analyzed for DNA adduct formation by accelerator mass spectrometry. Wistar rats produced several PhIP and N-hydroxy-PhIP glucuronides that accounted for approximately 25% of the total amount of recovered urinary metabolites. In the Gunn rats, PhIP and N-hydroxy-PhIP glucuronides were reduced by 68-92%, compared with the Wistar rats. PhIP-DNA adduct analysis from the Gunn rats revealed a correlation between reduced urinary PhIP and N-hydroxy-PhIP glucuronide levels and increased hepatic DNA adducts, compared with the Wistar rats. In the colon, DNA adduct levels were lower in the Gunn rats compared with the Wistar rats, suggesting deficient hepatic UGT1A activity provides protection against DNA adduct formation in peripheral tissue. Due to differences in PhIP metabolism between humans and rodents, extrapolation of these results to the human situation must be done with caution. These results indicate that UGT1A-mediated glucuronidation of PhIP and N-hydroxy-PhIP is an important pathway for PhIP detoxification, and demonstrate the importance of tissue-specific metabolism. Tissues with reduced UGT1A activity can have a higher rate of PhIP activation and be more inclined to form DNA adducts compared with tissues with normal UGT1A activity.
Malfatti
Michael A
MA
Biosciences Directorate, Lawrence Livermore National Laboratory, Livermore, CA 94551, USA.
Ubick
Esther A
EA
Felton
James S
JS
D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D002851:Chromatography, High Pressure Liquid; D003106:Colon / Q000187:drug effects / Q000378:metabolism; D018736:DNA Adducts / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000378:metabolism*; D007093:Imidazoles / Q000633:toxicity* / Q000652:urine; D015151:Immunoblotting; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000633:toxicity / Q000652:urine; D051381:Rats; D011911:Rats, Gunn; D017208:Rats, Wistar; D019324:beta-Naphthoflavone / Q000008:administration & dosage; D000099148:UGT1A1 Enzyme
D002273:Carcinogens; D018736:DNA Adducts; D004791:Enzyme Inhibitors; D020719:Glucuronides; D014453:Glucuronosyltransferase; D007093:Imidazoles; D011725:Pyridines; D019324:beta-Naphthoflavone; D000099148:UGT1A1 Enzyme; C060533:2-hydroxyamino-1-methyl-6-phenylimidazo(4,5-b)pyridine; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine