Ethanol-mediated transplacental induction of CYP2E1 in fetal rat liver.
We examined the potential for the widely consumed xenobiotic ethanol to transplacentally induce fetal rat CYP2E1. Throughout gestation, rat dams were fed a liquid diet containing 5% ethanol or two separate control diets. At 2 days before term, the dams were killed, and maternal and embryonic tissues were collected. Immunoblot analysis of microsomes from fetal liver, placenta and maternal brain revealed a band that comigrated with adult liver CYP2E1. The identity of the immunoreactive protein in placenta, brain and fetal liver was substantiated as CYP2E1 through restriction enzyme digestion of a reverse transcription-polymerase chain reaction product. Quantification of immunoblots containing microsomes from maternal and fetal liver of ethanol-treated dams displayed a 1.4- and 2.4-fold increase in CYP2E1, respectively, compared with microsomes from pair-fed controls. Chlorzoxazone and low substrate concentrations of N-nitrosodimethylamine were used as metabolic probes for CYP2E1. The rate of chlorzoxazone metabolism by maternal hepatic microsomes from dams fed the 5% ethanol diet was 2.6-fold greater than that of controls. Conversely, a negligible increase was observed in the rate of metabolism by hepatic microsomes from ethanol-exposed fetuses compared with pair-fed animals. When N-nitrosodimethylamine demethylation was examined, these same fetal samples exhibited greater rates of activity (1.5-fold) compared with microsomes from control animals. However, this increase was not as great as expected considering the 2.4-fold increase in CYP2E1 protein. Collectively, fetuses exposed to a 5% ethanol diet throughout gestation exhibited transplacental induction of an hepatic CYP2E1 that may possess different catalytic properties from the analogous adult enzyme.
Carpenter
S P
SP
University of New Mexico, College of Pharmacy, Albuquerque, USA. scarpen@agi.org
Savage
D D
DD
Schultz
E D
ED
Raucy
J L
JL
D000818:Animals; D002384:Catalysis; D002753:Chlorzoxazone / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis* / Q000378:metabolism; D004790:Enzyme Induction; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D005333:Fetus / Q000201:enzymology; D007150:Immunohistochemistry; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology; D009603:Nitroso Compounds / Q000378:metabolism; D010920:Placenta / Q000187:drug effects*; D011247:Pregnancy; D051381:Rats
D009603:Nitroso Compounds; C008651:4-nitrosodimethylaniline; D000431:Ethanol; D019392:Cytochrome P-450 CYP2E1; D002753:Chlorzoxazone