title: Diminished acyl-CoA synthetase isoform 4 activity in INS 832/13 cells reduces cellular epoxyeicosatrienoic acid levels and results in impaired glucose-stimulated insulin secretion.
abstract: Glucose-stimulated insulin secretion (GSIS) in pancreatic beta-cells is potentiated by fatty acids (FA). The initial step in the metabolism of intracellular FA is the conversion to acyl-CoA by long chain acyl-CoA synthetases (Acsls). Because the predominantly expressed Acsl isoforms in INS 832/13 cells are Acsl4 and -5, we characterized the role of these Acsls in beta-cell function by using siRNA to knock down Acsl4 or Acsl5. Compared with control cells, an 80% suppression of Acsl4 decreased GSIS and FA-potentiated GSIS by 32 and 54%, respectively. Knockdown of Acsl5 did not alter GSIS. Acsl4 knockdown did not alter FA oxidation or long chain acyl-CoA levels. With Acsl4 knockdown, incubation with 17 mm glucose increased media epoxyeicosatrienoic acids (EETs) and reduced cell membrane levels of EETs. Further, exogenous EETs reduced GSIS in INS 832/13 cells, and in Acsl4 knockdown cells, an EET receptor antagonist partially rescued GSIS. These results strongly suggest that Acsl4 activates EETs to form EET-CoAs that are incorporated into glycerophospholipids, thereby sequestering EETs. Exposing INS 832/13 cells to arachidonate or linoleate reduced Acsl4 mRNA and protein expression and reduced GSIS. These data indicate that Acsl4 modulates GSIS by regulating the levels of unesterified EETs and that arachidonate controls the expression of its activator Acsl4.
authors.affiliation: Department of Medicine, Division of Endocrinology, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599-7172, USA. eklett@med.unc.edu
authors.forename: Eric L
authors.initials: EL
authors.lastname: Klett
authors.forename: Shufen
authors.initials: S
authors.lastname: Chen
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Edin
authors.forename: Lei O
authors.initials: LO
authors.lastname: Li
authors.forename: Olga
authors.initials: O
authors.lastname: Ilkayeva
authors.forename: Darryl C
authors.initials: DC
authors.lastname: Zeldin
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Newgard
authors.forename: Rosalind A
authors.initials: RA
authors.lastname: Coleman
mesh_terms: D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003066:Coenzyme A Ligases / Q000235:genetics / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology*; D020404:Glycerophospholipids / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D007340:Insulinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008563:Membrane Lipids / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D034622:RNA Interference; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000214:Acyl Coenzyme A; D005227:Fatty Acids; D020404:Glycerophospholipids; D007328:Insulin; D008563:Membrane Lipids; D024101:Mitochondrial Proteins; C050715:8,9-epoxyeicosatrienoic acid; C500958:Acsl4 protein, rat; D003066:Coenzyme A Ligases; C508147:Acsl5 protein, rat; D015126:8,11,14-Eicosatrienoic Acid; D005947:Glucose
keywords: Acyl-CoA Synthetase; Arachidonic Acid; Beta-Cell; Eicosanoid; Epoxyeicosatrienoic Acids; Insulin Secretion; Polyunsaturated Fatty Acids