O2-dependent hepatotoxicity due to ethylhexanol in the perfused rat liver: mitochondria as a site of action.
Toxicity of 2-ethylhexanol, a metabolite of diethylhexyl phthalate, was assessed in the perfused rat liver. Livers from starved rats were perfused with ethylhexanol (3 mM) dissolved in Krebs-Henseleit buffer (pH 7.4, 37 degrees C) saturated with 95% O2-5% CO2 in both the anterograde and retrograde direction. Following infusion of ethylhexanol, O2 uptake and ketone body formation were diminished by 50 and 80%, respectively, and cell damage, as assessed by the appearance of lactate dehydrogenase in the effluent perfusate, was apparent. Both inhibition of O2 uptake by ethylhexanol and the appearance of lactate dehydrogenase in the perfusate were dose-dependent. Only O2-rich upstream regions of the liver lobule were damaged as reflected by trypan blue uptake. Inhibition of O2 uptake by ethylhexanol was also reflected by a 60% decrease in the ATP/ADP ratio. Local rates of O2 uptake, measured using miniature electrodes placed on the liver surface, indicated that ethylhexanol only diminished O2 uptake in O2-rich upstream regions of the liver lobule regardless of the direction of flow. This phenomenon apparently can be explained by a direct effect of ethylhexanol on mitochondria in upstream regions since active state 3 rates of respiration were inhibited by ethylhexanol in isolated mitochondria. Ethylhexanol also caused a dose-dependent decrease in the mitochondrial membrane potential and an increase in the beta-hydroxybutyrate/acetoacetate (B/A) ratio. However, infusion of radical scavengers such as allopurinol, cianidanol and uric acid did not alter lactate dehydrogenase release due to ethylhexanol. Thus, the toxicity of ethylhexanol in the liver is dependent on local O2 tension and mitochondrial are primary targets.(ABSTRACT TRUNCATED AT 250 WORDS)
Keller
B J
BJ
Department of Pharmacology, University of North Carolina, Chapel Hill.
Yamanaka
H
H
Liang
D C
DC
Kauffman
F C
FC
Thurman
R G
RG
D000227:Adenine Nucleotides / Q000378:metabolism; D000818:Animals; D005260:Female; D005508:Food Deprivation; D000441:Hexanols / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D010100:Oxygen / Q000378:metabolism / Q000493:pharmacokinetics*; D051381:Rats; D011919:Rats, Inbred Strains
D000227:Adenine Nucleotides; D000441:Hexanols; D007770:L-Lactate Dehydrogenase; D010100:Oxygen; C034017:2-ethylhexanol