Evidence for the generation of transaminase inhibitor(s) during ethanol metabolism by rat liver homogenates: a potential mechanism for alcohol toxicity.
Since ethanol consumption decreases hepatic aminotransferase activities in vivo, mechanisms of ethanol-mediated transaminase inhibition were explored in vitro using mitochondria-depleted rat liver homogenates. When homogenates were incubated at 37 degrees with 50 mM ethanol for 1 hr, alanine aminotransferase decreased by 20%, while aspartate aminotransferase was unchanged. After 2 hr, aspartate aminotransferase decreased by 20% and by 3 hr, alanine and aspartate aminotransferases were decreased by 31 and 23%, respectively. Levels of acetaldehyde generated during ethanol oxidation were 525 +/- 47 microM at 1 hr, 855 +/- 14 microM at 2 hr, and 1293 +/- 140 microM at 3 hr. Although inhibition of alcohol oxidation with methylpyrazole or cyanide markedly decreased ethanol-mediated transaminase inhibition, neither incubation with acetate nor generation of reducing equivalents by oxidation of lactate, malate, xylitol, or sorbitol altered the activity of either enzyme. However, semicarbazide, an aldehyde scavenger, prevented inhibition of both aminotransferases by ethanol. Moreover, incubation with 5 mM acetaldehyde for 1 hr inhibited alanine and aspartate aminotransferases by 36 and 26%, respectively. Cyanamide, an aldehyde dehydrogenase inhibitor, had little effect on ethanol-mediated transaminase inhibition. Thus, metabolism of ethanol by rat liver homogenates produces transaminase inhibition similar to that described in vivo and this effect requires acetaldehyde generation but not acetaldehyde oxidation. Since addition of pyridoxal 5'-phosphate to assay mixes did not reverse ethanol effects, aminotransferase inhibition does not result from displacement of vitamin B6 coenzymes.
Solomon
L R
LR
Department of Medicine, Veterans Administration Medical Center, West Haven, Connecticut 06516.
D000079:Acetaldehyde / Q000494:pharmacology; D000410:Alanine Transaminase / Q000037:antagonists & inhibitors*; D000818:Animals; D001219:Aspartate Aminotransferases / Q000037:antagonists & inhibitors*; D003484:Cyanamide / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D000431:Ethanol / Q000378:metabolism* / Q000633:toxicity; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D011919:Rats, Inbred Strains; D012663:Semicarbazides / Q000494:pharmacology; D012966:Sodium Cyanide / Q000494:pharmacology; D013012:Sorbitol / Q000494:pharmacology
D012663:Semicarbazides; D000431:Ethanol; D003484:Cyanamide; D013012:Sorbitol; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D000079:Acetaldehyde; D012966:Sodium Cyanide