Mechanism of the synergistic induction of CYP2H by isopentanol plus ethanol: comparison to glutethimide and relation to induction of 5-aminolevulinate synthase.
We had previously found that combined treatment with isopentanol and ethanol synergistically induced CYP2H protein and activity in cultured chick nepatoytes. Here we investigated the mechanism of induction of CYP2H by the alcohols and whether they caused a coordinate induction of 5-aminolevulinate synthase (ALAS) mRNA. Treatment with isopentanol alone or in combination with ethanol resulted in coordinate increases in CYP2H1 and ALAS mRNAs. With isopentanol alone, the amounts of CYP2H1 and ALAS mRNAs at 4 to 6 h were similar to those observed after treatment with the alcohol combination, but declined by 11 h. Readdition of isopentanol at 11 h again increased the expression of both mRNAs, indicating that the decreases at 11 h were due to limiting amounts of inducer. Similar results were observed in cells exposed to low concentrations of glutethimide. In the combined alcohol treatment, increases in CYP2H1 and ALAS mRNAs were sustained from 4 h to 11 h after addition of the alcohols, but decreased to control levels by 24 h. Using pulse labeling to measure de novo synthesis of CYP2H1/2 protein, we found that the increases in CYP2H1/2 protein reflected the increases in CYP2H1 mRNA. The half-life of CYP2H1/2 protein, measured from pulse-chase experiments, was approximately twofold greater than the half-life of CYP2H1 mRNA. Our results indicate that the alcohols and glutethimide coordinately increase ALAS and CYP2H1 mRNA, and that increases in CYP2H1/2 protein arise from increases in its mRNA.
Louis
C A
CA
Veterans Administration Medical Center, White River Junction, Vermont, 05009, USA.
Wood
S G
SG
Walton
H S
HS
Sinclair
P R
PR
Sinclair
J F
JF
D000624:5-Aminolevulinate Synthetase / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D004790:Enzyme Induction / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D005984:Glutethimide / Q000378:metabolism / Q000494:pharmacology*; D006207:Half-Life; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D000439:Pentanols / Q000378:metabolism / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D013997:Time Factors
D000439:Pentanols; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D005984:Glutethimide; C029683:isopentyl alcohol; D000624:5-Aminolevulinate Synthetase