title: Insulin protects pancreatic acinar cells from palmitoleic acid-induced cellular injury.
abstract: Acute pancreatitis is a serious and sometimes fatal inflammatory disease where the pancreas digests itself. The non-oxidative ethanol metabolites palmitoleic acid (POA) and POA-ethylester (POAEE) are reported to induce pancreatitis caused by impaired mitochondrial metabolism, cytosolic Ca(2+) ([Ca(2+)]i) overload and necrosis of pancreatic acinar cells. Metabolism and [Ca(2+)]i are linked critically by the ATP-driven plasma membrane Ca(2+)-ATPase (PMCA) important for maintaining low resting [Ca(2+)]i. The aim of the current study was to test the protective effects of insulin on cellular injury induced by the pancreatitis-inducing agents, ethanol, POA, and POAEE. Rat pancreatic acinar cells were isolated by collagenase digestion and [Ca(2+)]i was measured by fura-2 imaging. An in situ [Ca(2+)]i clearance assay was used to assess PMCA activity. Magnesium green (MgGreen) and a luciferase-based ATP kit were used to assess cellular ATP depletion. Ethanol (100 mM) and POAEE (100 μM) induced a small but irreversible Ca(2+) overload response but had no significant effect on PMCA activity. POA (50-100 μM) induced a robust Ca(2+) overload, ATP depletion, inhibited PMCA activity, and consequently induced necrosis. Insulin pretreatment (100 nm for 30 min) prevented the POA-induced Ca(2+) overload, ATP depletion, inhibition of the PMCA, and necrosis. Moreover, the insulin-mediated protection of the POA-induced Ca(2+) overload was partially prevented by the phosphoinositide-3-kinase (PI3K) inhibitor, LY294002. These data provide the first evidence that insulin directly protects pancreatic acinar cell injury induced by bona fide pancreatitis-inducing agents, such as POA. This may have important therapeutic implications for the treatment of pancreatitis.
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: Aysha
authors.initials: A
authors.lastname: Samad
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: Andrew
authors.initials: A
authors.lastname: James
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: James
authors.initials: J
authors.lastname: Wong
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: Parini
authors.initials: P
authors.lastname: Mankad
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: John
authors.initials: J
authors.lastname: Whitehouse
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: Waseema
authors.initials: W
authors.lastname: Patel
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and.
authors.forename: Marta
authors.initials: M
authors.lastname: Alves-Simoes
authors.affiliation: the Hepatobiliary Surgery Unit, Manchester Royal Infirmary, M13 9WL Manchester, United Kingdom.
authors.forename: Ajith K
authors.initials: AK
authors.lastname: Siriwardena
authors.affiliation: From the Faculty of Life Sciences, The University of Manchester, M13 9NT Manchester and jason.bruce@manchester.ac.uk.
authors.forename: Jason I E
authors.initials: JI
authors.lastname: Bruce
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D016923:Cell Death; D002867:Chromones / Q000494:pharmacology; D000431:Ethanol / Q000008:administration & dosage / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D005453:Fluorescence; D007328:Insulin / Q000502:physiology*; D009025:Morpholines / Q000494:pharmacology; D010179:Pancreas / Q000166:cytology / Q000187:drug effects*; D053499:Plasma Membrane Calcium-Transporting ATPases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002867:Chromones; D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D007328:Insulin; D009025:Morpholines; C008757:palmitoleic acid; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D000431:Ethanol; D000255:Adenosine Triphosphate; D053499:Plasma Membrane Calcium-Transporting ATPases; D002118:Calcium
keywords: Calcium; Calcium ATPase; Calcium Transport; Insulin; PMCA; Pancreas; Pancreatitis