title: Eicosapentaenoic acid membrane incorporation impairs ABCA1-dependent cholesterol efflux via a protein kinase A signaling pathway in primary human macrophages.
abstract: A diet rich in n-3/n-6 polyunsaturated fatty acids (PUFAs) is cardioprotective. Dietary PUFAs affect the cellular phospholipids composition, which may influence the function of membrane proteins. We investigated the impact of the membrane incorporation of several PUFAs on ABCA1-mediated cholesterol efflux, a key antiatherogenic pathway. Arachidonic acid (AA) (C20:4 n-6) and docosahexaenoic acid (DHA) (C22:6 n-3) decreased or increased cholesterol efflux from J774 mouse macrophages, respectively, whereas they had no effect on efflux from human monocyte-derived macrophages (HMDM). Importantly, eicosapentaenoic acid (EPA) (C20:5 n-3) induced a dose-dependent reduction of ABCA1 functionality in both cellular models (-28% for 70μM of EPA in HMDM), without any alterations in ABCA1 expression. These results show that PUFA membrane incorporation does not have the same consequences on cholesterol efflux from mouse and human macrophages. The EPA-treated HMDM exhibited strong phospholipid composition changes, with high levels of both EPA and its elongation product docosapentaenoic acid (DPA) (C22:5 n-3), which is associated with a decreased level of AA. In HMDM, EPA reduced the ATPase activity of the membrane transporter. Moreover, the activation of adenylate cyclase by forskolin and the inhibition of cAMP phosphodiesterase by isobutylmethylxanthine restored ABCA1 cholesterol efflux in EPA-treated human macrophages. In conclusion, EPA membrane incorporation reduces ABCA1 functionality in mouse macrophages as well as in primary human macrophages and this effect seems to be PKA-dependent in human macrophages.
authors.affiliation: Lip(Sys) 2, Athérosclérose: homéostasie et trafic du cholestérol des macrophages,(1) Univ Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France; Laboratoire de Biochimie, AP-HP (Assistance Publique-Hôpitaux de Paris), Hôpital Européen Georges Pompidou, 75015 Paris, France. Electronic address: natalie_fournier@yahoo.fr.
authors.forename: Natalie
authors.initials: N
authors.lastname: Fournier
authors.affiliation: Lip(Sys) 2, Athérosclérose: homéostasie et trafic du cholestérol des macrophages,(1) Univ Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Tardivel
authors.affiliation: Laboratoire de Biochimie hormonale, AP-HP (Assistance Publique-Hôpitaux de Paris), Hôpital Robert Debré, 75019 Paris, France.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Benoist
authors.affiliation: Laboratoire de Biochimie, AP-HP (Assistance Publique-Hôpitaux de Paris), Hôpital Européen Georges Pompidou, 75015 Paris, France.
authors.forename: Benoît
authors.initials: B
authors.lastname: Vedie
authors.affiliation: UMR INRA 1198, Biologie du Développement et Reproduction, Jouy-en-Josas, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Rousseau-Ralliard
authors.affiliation: Lip(Sys) 2, Athérosclérose: homéostasie et trafic du cholestérol des macrophages,(1) Univ Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Nowak
authors.affiliation: Lip(Sys) 2, Athérosclérose: homéostasie et trafic du cholestérol des macrophages,(1) Univ Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France.
authors.forename: Fatima
authors.initials: F
authors.lastname: Allaoui
authors.affiliation: Lip(Sys) 2, Athérosclérose: homéostasie et trafic du cholestérol des macrophages,(1) Univ Paris-Sud, Université Paris-Saclay, 92296 Châtenay-Malabry, France; Laboratoire de Biochimie, AP-HP (Assistance Publique-Hôpitaux de Paris), Hôpital Européen Georges Pompidou, 75015 Paris, France.
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Paul
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism; D064286:ATP Binding Cassette Transporter 1 / Q000235:genetics / Q000378:metabolism*; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology; D001692:Biological Transport; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004281:Docosahexaenoic Acids / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D015118:Eicosapentaenoic Acid / Q000378:metabolism / Q000494:pharmacology*; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D061251:Primary Cell Culture; D000067996:RAW 264.7 Cells; D015398:Signal Transduction / Q000187:drug effects*; D013045:Species Specificity
chemical_list: D015105:3',5'-Cyclic-AMP Phosphodiesterases; D064286:ATP Binding Cassette Transporter 1; D000262:Adenylyl Cyclases; D016718:Arachidonic Acid; D002784:Cholesterol; D017868:Cyclic AMP-Dependent Protein Kinases; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid; D005231:Fatty Acids, Unsaturated; C577836:ABCA1 protein, human; C577833:Abca1 protein, mouse; C026219:docosapentaenoic acid
keywords: ABCA1-mediated cholesterol efflux; EPA; Human monocyte-derived macrophages; J774 mouse macrophages; Membrane fatty acid composition; PKA