Differences between human and rat intestinal and hepatic bisphenol A glucuronidation and the influence of alamethicin on in vitro kinetic measurements.
The extent to which membrane-disrupting agents, such as alamethicin, may alter cofactor transport and influence in vitro kinetic measurements of glucuronidation is a major concern regarding the characterization and extrapolation of inter- and intraspecies pharmacokinetics of bisphenol A (BPA). An additional concern is the omission of a BPA intestinal metabolism component in current pharmacokinetic models used to assess oral exposure. In this study, BPA glucuronidation in native hepatic microsomes from female rat and female human liver displayed higher V(max) values than that in males. In the presence of alamethicin, all hepatic V(max) values increased; however, this increase was disproportionately greater in males and gender differences were no longer observed. Female rats exhibited a much higher K(m) than all other species and genders; the addition of alamethicin had little influence on K(m) values for any of the test systems. The dissimilar K(m) measured for female rat suggests that different UDP-glucuronosyltransferase (UGT) enzyme(s) are involved in BPA glucuronidation. The presence of different UGTs in female rat was confirmed using Hill coefficients measured from diclofenac-mediated chemical inhibition assays within hepatic microsomes and purified human UGT2B7 and UGT2B15. Mixed-gender human intestinal microsomes showed little BPA glucuronidation reactivity compared with those from male rat intestine. Male rat intestinal microsomes in the presence of alamethicin exhibited a V(max) that was nearly 30-fold higher than that for mixed human microsomes. The species and gender metabolic differences we observed between rat and human liver and intestine provide key information for delineating BPA pharmacokinetics needed for human health risk assessment.
Mazur
Christopher S
CS
U.S. Environmental Protection Agency, National Exposure Research Laboratory, Ecosystems Research Division, Athens, GA 30605, USA. mazur.chris@epa.gov
Kenneke
John F
JF
Hess-Wilson
Janet K
JK
Lipscomb
John C
JC
D000408:Alamethicin / Q000494:pharmacology*; D000818:Animals; D001559:Benzhydryl Compounds; D004347:Drug Interactions; D005260:Female; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000502:physiology; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008297:Male; D010636:Phenols / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012727:Sex Characteristics; D013045:Species Specificity; D000098826:Bisphenol A Compounds
D000408:Alamethicin; D001559:Benzhydryl Compounds; D020719:Glucuronides; D014453:Glucuronosyltransferase; D010636:Phenols; C491474:UGT2B7 protein, human; C472000:UDP-glucuronosyltransferase 2B15, human; C006780:bisphenol A; D000098826:Bisphenol A Compounds