title: Fatty acid ethyl ester synthase inhibition ameliorates ethanol-induced Ca2+-dependent mitochondrial dysfunction and acute pancreatitis.
abstract: OBJECTIVE
  Non-oxidative metabolism of ethanol (NOME) produces fatty acid ethyl esters (FAEEs) via carboxylester lipase (CEL) and other enzyme action implicated in mitochondrial injury and acute pancreatitis (AP). This study investigated the relative importance of oxidative and non-oxidative pathways in mitochondrial dysfunction, pancreatic damage and development of alcoholic AP, and whether deleterious effects of NOME are preventable.
  
  
  METHODS
  Intracellular calcium ([Ca(2+)](C)), NAD(P)H, mitochondrial membrane potential and activation of apoptotic and necrotic cell death pathways were examined in isolated pancreatic acinar cells in response to ethanol and/or palmitoleic acid (POA) in the presence or absence of 4-methylpyrazole (4-MP) to inhibit oxidative metabolism. A novel in vivo model of alcoholic AP induced by intraperitoneal administration of ethanol and POA was developed to assess the effects of manipulating alcohol metabolism.
  
  
  RESULTS
  Inhibition of OME with 4-MP converted predominantly transient [Ca(2+)](C) rises induced by low ethanol/POA combination to sustained elevations, with concurrent mitochondrial depolarisation, fall of NAD(P)H and cellular necrosis in vitro. All effects were prevented by 3-benzyl-6-chloro-2-pyrone (3-BCP), a CEL inhibitor. 3-BCP also significantly inhibited rises of pancreatic FAEE in vivo and ameliorated acute pancreatic damage and inflammation induced by administration of ethanol and POA to mice.
  
  
  CONCLUSIONS
  A combination of low ethanol and fatty acid that did not exert deleterious effects per se became toxic when oxidative metabolism was inhibited. The in vitro and in vivo damage was markedly inhibited by blockade of CEL, indicating the potential for development of specific therapy for treatment of alcoholic AP via inhibition of FAEE generation.
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK Department of Integrated Traditional Chinese and Western Medicine, Sichuan Provincial Pancreatitis Centre, West China Hospital, Sichuan University, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: David M
authors.initials: DM
authors.lastname: Booth
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Cane
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Chvanov
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Muhammad A
authors.initials: MA
authors.lastname: Javed
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Elliott
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Jane A
authors.initials: JA
authors.lastname: Armstrong
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Hayley
authors.initials: H
authors.lastname: Dingsdale
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Nicole
authors.initials: N
authors.lastname: Cash
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: William
authors.initials: W
authors.lastname: Greenhalf
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Rajarshi
authors.initials: R
authors.lastname: Mukherjee
authors.affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, Texas, USA.
authors.forename: Bhupendra S
authors.initials: BS
authors.lastname: Kaphalia
authors.affiliation: Morvus Technology Limited, Carmarthen, UK.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Jaffar
authors.affiliation: Cardiff School of Biosciences, University of Cardiff, Cardiff, UK.
authors.forename: Ole H
authors.initials: OH
authors.lastname: Petersen
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Alexei V
authors.initials: AV
authors.lastname: Tepikin
authors.affiliation: NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: Robert
authors.initials: R
authors.lastname: Sutton
authors.affiliation: Department of Cellular & Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK NIHR Liverpool Pancreas Biomedical Research Unit, RLBUHT, Institute of Translational Medicine, University of Liverpool, Liverpool, Merseyside, UK.
authors.forename: David N
authors.initials: DN
authors.lastname: Criddle
mesh_terms: D061354:Acinar Cells / Q000187:drug effects / Q000378:metabolism; D000217:Acyltransferases / Q000037:antagonists & inhibitors*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D043182:Carboxylesterase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D000431:Ethanol / Q000378:metabolism* / Q000633:toxicity; D005227:Fatty Acids / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology; D000077604:Fomepizole; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D009336:Necrosis; D019512:Pancreatitis, Alcoholic / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D011720:Pyrazoles / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology*
chemical_list: D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D011720:Pyrazoles; D011753:Pyrones; C008757:palmitoleic acid; D000431:Ethanol; D009249:NADP; D000077604:Fomepizole; C037903:3-benzyl-6-chloro-2-pyrone; D000217:Acyltransferases; D043182:Carboxylesterase; C043403:fatty acyl ethyl ester synthase; D002118:Calcium
keywords: ACUTE Pancreatitis; Alcohol-Induced Injury; Calcium; Ethanol; Pancreatic Damage