title: Comparison of acetaminophen toxicity in primary hepatocytes isolated from transgenic mice with different appolipoprotein E alleles.
abstract: The nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor, important for combating electrophilic and oxidative stress in the liver and other organs. This encompasses detoxification of hepatotoxic drugs, including acetaminophen (APAP). Recently, an association between apolipoprotein E (ApoE) genotype and Nrf2 expression was described. We compared the toxicity of APAP on primary culture hepatocytes isolated from transgenic mice carrying two different human ApoE alleles and wild-type controls. The cells were exposed to APAP in concentrations from 0.5 to 4 mM for up to 24 hours. APAP led to a dose-dependent hepatotoxicity from 1 mM after 16 h exposure in all mice tested. The toxicity was higher in hepatocytes isolated from both transgenic strains than in wild-type controls and most pronounced in ApoE3 mice. Concurrently, there was a decline in mitochondrial membrane potential, especially in ApoE3 hepatocytes. The formation of reactive oxygen species was increased after 24 hours with 2.5 mM APAP in hepatocytes of all strains tested, with the highest increase being in the ApoE3 genotype. The activity of caspases 3 and 7 did not differ among groups and was minimal after 24 hour incubation with 4 mM APAP. We observed higher lipid accumulation in hepatocytes isolated from both transgenic strains than in wild-type controls. The expression of Nrf2-dependent genes was higher in ApoE3 than in ApoE4 hepatocytes and some of these genes were induced by APAP treatment. In conclusion, transgenic mice with ApoE4 and ApoE3 alleles displayed higher susceptibility to acute APAP toxicity in vitro than wild-type mice. Of the two transgenic genotypes tested, ApoE3 allele carriers were more prone to injury.
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic. mezev5ar@lfhk.cuni.cz.
authors.forename: V
authors.initials: V
authors.lastname: Mezera
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: O
authors.initials: O
authors.lastname: Kucera
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: A
authors.initials: A
authors.lastname: Moravcova
authors.affiliation: Department of Medical Biochemistry, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: E
authors.initials: E
authors.lastname: Peterova
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: T
authors.initials: T
authors.lastname: Rousar
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: D
authors.initials: D
authors.lastname: Rychtrmoc
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: O
authors.initials: O
authors.lastname: Sobotka
authors.affiliation: Department of Physiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague, Hradec Kralove, Czech Republic.
authors.forename: Z
authors.initials: Z
authors.lastname: Cervinkova
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D000483:Alleles; D018712:Analgesics, Non-Narcotic / Q000633:toxicity*; D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics*; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D005838:Genotype; D005978:Glutathione / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D050356:Lipid Metabolism / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D051267:NF-E2-Related Factor 2 / Q000235:genetics; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D018712:Analgesics, Non-Narcotic; D001057:Apolipoproteins E; D051267:NF-E2-Related Factor 2; D017382:Reactive Oxygen Species; D000082:Acetaminophen; D005978:Glutathione