title: N-acylethanolamine-hydrolyzing acid amidase and fatty acid amide hydrolase inhibition differentially affect N-acylethanolamine levels and macrophage activation.
abstract: N-acylethanolamines (NAEs) such as N-palmitoylethanolamine and anandamide are endogenous bioactive lipids having numerous functions, including the control of inflammation. Their levels and therefore actions can be controlled by modulating the activity of two hydrolytic enzymes, N-acylethanolamine-hydrolyzing acid amidase (NAAA) and fatty acid amide hydrolase (FAAH). As macrophages are key to inflammatory processes, we used lipopolysaccharide-activated J774 macrophages, as well as primary mouse alveolar macrophages, to study the effect of FAAH and NAAA inhibition, using PF-3845 and AM9053 respectively, on macrophage activation and NAE levels measured by HPLC-MS. Markers of macrophage activation were measured by qRT-PCR and ELISA. Activation of macrophages decreased NAAA expression and NAE hydrolytic activity. FAAH and NAAA inhibition increased the levels of the different NAEs, although with different magnitudes, whether in control condition or following LPS-induced macrophage activation. Both inhibitors reduced several markers of macrophage activation, such as mRNA expression of inflammatory mediators, as well as cytokine and prostaglandin production, with however some differences between FAAH and NAAA inhibition. Most of the NAEs tested - including N-docosatetraenoylethanolamine and N-docosahexaenoylethanolamine - also reduced LPS-induced mRNA expression of inflammatory mediators, again with differences depending on the marker and the NAE, thus offering a potential explanation for the differential effect of the inhibitors on macrophage activation markers. In conclusion, we show different and complementary effects of NAE on lipopolysaccharide-induced macrophage activation. Our results support an important role for inhibition of NAE hydrolysis and NAAA inhibition in particular in controlling macrophage activation, and thus inflammation.
authors.affiliation: Bioanalysis and Pharmacology of Bioactive Lipids Research Group, Louvain Drug Research Institute, Université Catholique de Louvain, Bruxelles, Belgium.
authors.forename: Mireille
authors.initials: M
authors.lastname: Alhouayek
authors.affiliation: Bioanalysis and Pharmacology of Bioactive Lipids Research Group, Louvain Drug Research Institute, Université Catholique de Louvain, Bruxelles, Belgium.
authors.forename: Pauline
authors.initials: P
authors.lastname: Bottemanne
authors.affiliation: Center for Drug Discovery, Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, United States; Department of Pharmaceutical Sciences, Northeastern University, Boston, MA 02115, United States.
authors.forename: Alexandros
authors.initials: A
authors.lastname: Makriyannis
authors.affiliation: Bioanalysis and Pharmacology of Bioactive Lipids Research Group, Louvain Drug Research Institute, Université Catholique de Louvain, Bruxelles, Belgium. Electronic address: giulio.muccioli@uclouvain.be.
authors.forename: Giulio G
authors.initials: GG
authors.lastname: Muccioli
mesh_terms: D000577:Amides; D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism; D063388:Endocannabinoids / Q000378:metabolism; D004983:Ethanolamines / Q000737:chemistry / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D007249:Inflammation / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000633:toxicity; D008262:Macrophage Activation / Q000187:drug effects / Q000235:genetics; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D010169:Palmitic Acids / Q000737:chemistry / Q000378:metabolism*; D010880:Piperidines / Q000008:administration & dosage; D053284:Polyunsaturated Alkamides / Q000378:metabolism; D011725:Pyridines / Q000008:administration & dosage; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000577:Amides; D000581:Amidohydrolases; D001095:Arachidonic Acids; D063388:Endocannabinoids; D004983:Ethanolamines; D008070:Lipopolysaccharides; D010169:Palmitic Acids; D010880:Piperidines; D053284:Polyunsaturated Alkamides; D011725:Pyridines; D000099252:Fatty Acid Amide Hydrolases; C022203:N-acylethanolamines; C557973:PF 3845; C005958:palmidrol; C582821:NAAA protein, mouse; C078814:anandamide
keywords: DHEA, DTEA; Docosahexaenoylethanolamide; Docosatetraenoylethanolamide; PEA; Palmitoylethanolamide