title: A cellular model to study drug-induced liver injury in nonalcoholic fatty liver disease: Application to acetaminophen.
abstract: Obesity and nonalcoholic fatty liver disease (NAFLD) can increase susceptibility to hepatotoxicity induced by some xenobiotics including drugs, but the involved mechanisms are poorly understood. For acetaminophen (APAP), a role of hepatic cytochrome P450 2E1 (CYP2E1) is suspected since the activity of this enzyme is consistently enhanced during NAFLD. The first aim of our study was to set up a cellular model of NAFLD characterized not only by triglyceride accumulation but also by higher CYP2E1 activity. To this end, human HepaRG cells were incubated for one week with stearic acid or oleic acid, in the presence of different concentrations of insulin. Although cellular triglycerides and the expression of lipid-responsive genes were similar with both fatty acids, CYP2E1 activity was significantly increased only by stearic acid. CYP2E1 activity was reduced by insulin and this effect was reproduced in cultured primary human hepatocytes. Next, APAP cytotoxicity was assessed in HepaRG cells with or without lipid accretion and CYP2E1 induction. Experiments with a large range of APAP concentrations showed that the loss of ATP and glutathione was almost always greater in the presence of stearic acid. In cells pretreated with the CYP2E1 inhibitor chlormethiazole, recovery of ATP was significantly higher in the presence of stearate with low (2.5mM) or high (20mM) concentrations of APAP. Levels of APAP-glucuronide were significantly enhanced by insulin. Hence, HepaRG cells can be used as a valuable model of NAFLD to unveil important metabolic and hormonal factors which can increase susceptibility to drug-induced hepatotoxicity.
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France.
authors.forename: Anaïs
authors.initials: A
authors.lastname: Michaut
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France.
authors.forename: Dounia
authors.initials: D
authors.lastname: Le Guillou
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France; Service de Biochimie et Toxicologie, CHU Pontchaillou, Rennes, France.
authors.forename: Caroline
authors.initials: C
authors.lastname: Moreau
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France.
authors.forename: Simon
authors.initials: S
authors.lastname: Bucher
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Mitchell R
authors.initials: MR
authors.lastname: McGill
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Martinais
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France; Service de Biochimie et Toxicologie, CHU Pontchaillou, Rennes, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gicquel
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France; Service de Biochimie et Toxicologie, CHU Pontchaillou, Rennes, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Morel
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France.
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Robin
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, KS, USA.
authors.forename: Hartmut
authors.initials: H
authors.lastname: Jaeschke
authors.affiliation: INSERM, U991, Université de Rennes 1, Rennes, France. Electronic address: bernard.fromenty@inserm.fr.
authors.forename: Bernard
authors.initials: B
authors.lastname: Fromenty
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D002460:Cell Line; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism*; D065700:Cytochrome P-450 CYP2E1 Inducers / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D005227:Fatty Acids / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D065626:Non-alcoholic Fatty Liver Disease / Q000139:chemically induced* / Q000378:metabolism*
chemical_list: D065700:Cytochrome P-450 CYP2E1 Inducers; D005227:Fatty Acids; D000082:Acetaminophen
keywords: Acetaminophen; CYP2E1; Hepatotoxicity; Liver; NAFLD; Obesity