Use of lymphocytes for assessing ethanol-mediated alterations in the expression of hepatic cytochrome P4502E1.
The ethanol-inducible cytochrome P4502E1 (2E1) is involved in the bioactivation of numerous hepatotoxins and hepatocarcinogens. Because high levels of expression may enhance the degree and severity of hepatotoxicity from exposure to chemicals metabolized by this enzyme, a relatively noninvasive method to phenotypically distinguish those individuals exhibiting elevated concentrations of 2E1 may be useful. With this in mind, we examined whether ethanol exposure could alter 2E1 in rabbit white blood cells and liver in a similar manner. Microsomes prepared from freshly isolated, rather than cultured cells, were used to immunochemically detect 2E1. The enzyme was found in lymphocytes and neutrophils. Lymphocytes, which comprise the majority of the white cell population in rabbits, were monitored for changes in 2E1 protein levels after ethanol exposure and compared with alterations of the hepatic enzyme. Results presented herein demonstrate that the degree of enhancement in 2E1 expression of lymphocytes and liver was dependent on the length and dose of alcohol exposure. Indeed, correlations were observed between blood alcohol concentrations and 2E1 content in lymphocytes (r = 0.65, p < 0.01) and liver (r = 0.60, p < 0.01). The greatest increase in 2E1 (6- to 10-fold) occurred in both liver and lymphocytes at a dose of 15% ethanol for 12 days of treatment. This induction was evident regardless of whether blood was taken from treated and compared with untreated rabbits or if white cells were obtained from the same animal before and after ethanol exposure. The latter findings demonstrate that changes in lymphocyte 2E1 were caused by ethanol exposure and not to variability in enzyme expression among rabbits. Interestingly, at the 10% dose, elevation of 2E1 was noted as early as 3 days, declined at 6 days, and at 12 and 24 days returned to slightly higher levels than those seen at the 3-day exposure period. This pattern of 2E1 elevation was observed in both the liver and lymphocytes. In fact, at all exposure periods and at the two doses of alcohol examined, a correlation (r = 0.70, p < 0.01) was observed between lymphocyte and liver 2E1 content. Collectively, these studies show that induction of 2E1 in lymphocytes and liver occurs in a parallel fashion. Furthermore, results suggest that blood 2E1 may be used in humans as a phenotypic marker for xenobiotic-promoted alterations in the expression of the liver enzyme. These findings should have a significant impact on in vivo monitoring of this P450 enzyme.
Raucy
J L
JL
University of New Mexico, College of Pharmacy, Albuquerque, USA.
Curley
G
G
Carpenter
S P
SP
D000437:Alcoholism / Q000201:enzymology / Q000235:genetics*; D000818:Animals; D019392:Cytochrome P-450 CYP2E1; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004790:Enzyme Induction / Q000235:genetics; D000431:Ethanol / Q000493:pharmacokinetics*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005819:Genetic Markers / Q000235:genetics; D008214:Lymphocytes / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics*; D010641:Phenotype; D011817:Rabbits
D005819:Genetic Markers; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D019392:Cytochrome P-450 CYP2E1; D010089:Oxidoreductases, N-Demethylating