title: Enzymic catalysis of the accumulation of acetaldehyde from ethanol in human prenatal cephalic tissues: evaluation of the relative contributions of CYP2E1, alcohol dehydrogenase, and catalase/peroxidases.
abstract: BACKGROUND
  The human prenatal brain is very sensitive to the toxic effects of ethanol, but very little information is available concerning the conversion of ethanol to the highly cytotoxic metabolite, acetaldehyde, in that organ. Thus, experiments were designed to investigate rates of accumulation of acetaldehyde from ethanol in the prenatal human brain.
  
  
  METHODS
  Prenatal human cephalic tissue homogenates were used as enzyme sources and were compared with analogous preparations of adult rat livers. Generated acetaldehyde was derivatized with cyclohexane-1,3-dione to yield fluorescent decahydroacrizine-1,8-dione, which was readily separated, detected, and quantitated with HPLC.
  
  
  RESULTS
  Detected rates of accumulation were unexpectedly high, even in the absence of added NADPH, NAD+, or H2O2, which are cofactors/cosubstrates for cytochrome P-450-, alcohol dehydrogenase- and catalase/peroxidase-catalyzed reactions, respectively. Without added cofactors/cosubstrates or other components and under linear reaction conditions, rates in human prenatal cephalic preparations were approximately 20% of those observed with analogous preparations of adult rat livers. Cofactor/cosubstrate-independent reactions were localized in the cytosolic (soluble) fraction and were strongly dependent on molecular oxygen (O2). They were not inhibited substantially by carbon monoxide (CO:O2 = 80:20 vs N2:O2 = 80:20) or by pyrazole in concentrations up to 10 mM and were only weakly inhibited by azide. Preincubations with excess catalase did not result in decreased activity. Reactions exhibited substrate saturation and heat inactivation indicating enzymic catalysis.
  
  
  CONCLUSIONS
  Experiments indicated a relatively rapid accumulation of acetaldehyde from ethanol in human prenatal brain tissues and suggested that the observed cofactor/cosubstrate-independent reactions were largely independent of P-450 cytochromes, alcohol dehydrogenases, or catalase/peroxidases. Results were consistent with catalysis by an as yet unidentified cytosolic oxidase(s).
authors.affiliation: Department of Pharmacology, School of Medicine, University of Washington, Seattle 98195, USA.
authors.forename: R E
authors.initials: RE
authors.lastname: Person
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.forename: A G
authors.initials: AG
authors.lastname: Fantel
authors.forename: M R
authors.initials: MR
authors.lastname: Juchau
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000196:embryology / Q000201:enzymology*; D002374:Catalase / Q000378:metabolism; D002492:Central Nervous System Depressants / Q000378:metabolism*; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008297:Male; D010544:Peroxidases / Q000378:metabolism; D051381:Rats
chemical_list: D002492:Central Nervous System Depressants; D000431:Ethanol; D000426:Alcohol Dehydrogenase; D010544:Peroxidases; D002374:Catalase; D019392:Cytochrome P-450 CYP2E1; D000079:Acetaldehyde