title: Low-dose acetaminophen induces early disruption of cell-cell tight junctions in human hepatic cells and mouse liver.
abstract: Dysfunction of cell-cell tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. Liver TJs preserve cellular polarity by delimiting functional bile-canalicular structures, forming the blood-biliary barrier. In acetaminophen-hepatotoxicity, the mechanism by which tissue cohesion and polarity are affected remains unclear. Here, we demonstrate that acetaminophen, even at low-dose, disrupts the integrity of TJ and cell-matrix adhesions, with indicators of cellular stress with liver injury in the human hepatic HepaRG cell line, and primary hepatocytes. In mouse liver, at human-equivalence (therapeutic) doses, dose-dependent loss of intercellular hepatic TJ-associated ZO-1 protein expression was evident with progressive clinical signs of liver injury. Temporal, dose-dependent and specific disruption of the TJ-associated ZO-1 and cytoskeletal-F-actin proteins, correlated with modulation of hepatic ultrastructure. Real-time impedance biosensing verified in vitro early, dose-dependent quantitative decreases in TJ and cell-substrate adhesions. Whereas treatment with NAPQI, the reactive metabolite of acetaminophen, or the PKCα-activator and TJ-disruptor phorbol-12-myristate-13-acetate, similarly reduced TJ integrity, which may implicate oxidative stress and the PKC pathway in TJ destabilization. These findings are relevant to the clinical presentation of acetaminophen-hepatotoxicity and may inform future mechanistic studies to identify specific molecular targets and pathways that may be altered in acetaminophen-induced hepatic depolarization.
authors.affiliation: MRC Centre for Regenerative Medicine, SCRM Building, The University of Edinburgh, Edinburgh BioQuarter, 5 Little France Drive, Edinburgh EH16 4UU, UK.
authors.forename: Wesam
authors.initials: W
authors.lastname: Gamal
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Philipp
authors.initials: P
authors.lastname: Treskes
authors.affiliation: Scottish National Blood Transfusion Service, Research, Development and Innovation Directorate, Cell Therapy Group, Ellens Glen Road, Edinburgh, EH17 7QT, UK.
authors.forename: Kay
authors.initials: K
authors.lastname: Samuel
authors.affiliation: Department of Biochemistry, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway, UK.|Norwegian Center for Stem Cell Research, PO Box 1112 Blindern, 0317 Oslo, Norway.|Institute of Immunology, Oslo University Hospital-Rikshospitalet, PO Box 4950 Nydalen, Oslo 0424, Norway.
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Sullivan
authors.affiliation: Department of Biochemistry, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway, UK.
authors.forename: Richard
authors.initials: R
authors.lastname: Siller
authors.affiliation: Institute for Bioengineering, University of Edinburgh, King's Buildings, Colin MacLaurin Road, EH9 3DW, UK.
authors.forename: Vlastimil
authors.initials: V
authors.lastname: Srsen
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Katie
authors.initials: K
authors.lastname: Morgan
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Bryans
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Ada
authors.initials: A
authors.lastname: Kozlowska
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Andreas
authors.initials: A
authors.lastname: Koulovasilopoulos
authors.affiliation: Institute for Integrated Micro and Nano systems, University of Edinburgh, Scottish Micro Electronic Centre, Alexander Crum Brown Road, EH9 3FF, UK.
authors.forename: Ian
authors.initials: I
authors.lastname: Underwood
authors.affiliation: Institute for Bioengineering, University of Edinburgh, King's Buildings, Colin MacLaurin Road, EH9 3DW, UK.
authors.forename: Stewart
authors.initials: S
authors.lastname: Smith
authors.affiliation: Easter Bush Pathology, The Royal (Dick) School of Veterinary Studies and The Roslin Institute, Easter Bush Campus, Midlothian, EH25 9RG, UK.
authors.forename: Jorge
authors.initials: J
authors.lastname: Del-Pozo
authors.affiliation: Easter Bush Pathology, The Royal (Dick) School of Veterinary Studies and The Roslin Institute, Easter Bush Campus, Midlothian, EH25 9RG, UK.
authors.forename: Sharon
authors.initials: S
authors.lastname: Moss
authors.affiliation: MRC Centre for Inflammation Research, The Queen's Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, UK.
authors.forename: Alexandra Inés
authors.initials: AI
authors.lastname: Thompson
authors.affiliation: MRC Centre for Inflammation Research, The Queen's Medical Research Institute, University of Edinburgh, Edinburgh EH16 4TJ, UK.
authors.forename: Neil C
authors.initials: NC
authors.lastname: Henderson
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Hayes
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: John N
authors.initials: JN
authors.lastname: Plevris
authors.affiliation: MRC Centre for Regenerative Medicine, SCRM Building, The University of Edinburgh, Edinburgh BioQuarter, 5 Little France Drive, Edinburgh EH16 4UU, UK.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Bagnaninchi
authors.affiliation: Hepatology Laboratory, University of Edinburgh, Royal Infirmary of Edinburgh, 49 Little France Crescent EH16 4SB, UK.
authors.forename: Leonard J
authors.initials: LJ
authors.lastname: Nelson
mesh_terms: D000082:Acetaminophen / Q000009:adverse effects*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology*; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress; D019108:Tight Junctions / Q000473:pathology*; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D000199:Actins; C569043:Tjp1 protein, mouse; D062826:Zonula Occludens-1 Protein; D000082:Acetaminophen