Anandamide inhibits IL-12p40 production by acting on the promoter repressor element GA-12: possible involvement of the COX-2 metabolite prostamide E(2).
The eCB [endoCB (cannabinoid)] system is being considered as a novel therapeutic target for immune disorders. Cytokines of the IL-12 (interleukin-12) family have essential functions in cell-mediated immunity. In the present study, we have addressed the mechanisms of action of the eCB AEA (anandamide) on the regulation of IL-12p40 in activated microglia/macrophages. We demonstrated that AEA can inhibit the expression of p35, p19 and p40 subunits, which form the biologically-active cytokines IL-12 and IL-23 in microglia stimulated with LPS (lipopolysaccharide)/IFNgamma (interferon gamma). Additionally, we have provided evidence that AEA reduces the transcriptional activity of the IL-12p40 gene in LPS- and IFNgamma-co-activated cells, and this is independent of CB or vanilloid receptor activation. Site-directed mutageneis of the different elements of the p40 promoter showed that AEA regulates IL-12p40 expression by acting on the repressor site GA-12 (GATA sequence in IL-12 promoter). Prostamide E(2) (prostaglandin E(2) ethanolamide), a product considered to be a putative metabolite of AEA by COX-2 (cyclo-oxygenase 2) oxygenation, was also able to inhibit the activity of the IL-12p40 promoter by acting at the repressor site. The effects of AEA and prostamide E(2) on p40 transcription were partially reversed by an antagonist of EP(2) (prostanoid receptor-type 2), suggesting the possibility that prostamide E(2) may contribute to the effects of AEA on IL-12p40 gene regulation. Accordingly, the inhibition of COX-2 by NS-398 partially reversed the inhibitory effects of AEA on IL-12 p40. Overall, our findings provide new mechanistic insights into the activities of AEA in immune-related disorders, which may be relevant for the clinical management of such diseases.
Correa
Fernando
F
Neuroimmunology Group, Functional and Systems Neurobiology Department, Instituto Cajal, CSIC (Consejo Superior de Investigaciones Científicas), Madrid, Spain.
Docagne
Fabian
F
Clemente
Diego
D
Mestre
Leyre
L
Becker
Christoph
C
Guaza
Carmen
C
D000818:Animals; D001095:Arachidonic Acids / Q000494:pharmacology*; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D015232:Dinoprostone / Q000031:analogs & derivatives* / Q000378:metabolism; D063388:Endocannabinoids; D005786:Gene Expression Regulation; D007371:Interferon-gamma / Q000378:metabolism; D053762:Interleukin-12 Subunit p40 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D053759:Interleukin-23 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D017628:Microglia / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D053284:Polyunsaturated Alkamides / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D021122:Protein Subunits / Q000378:metabolism; D043882:Receptors, Cannabinoid / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D001095:Arachidonic Acids; D063388:Endocannabinoids; D053762:Interleukin-12 Subunit p40; D053759:Interleukin-23; D008070:Lipopolysaccharides; D053284:Polyunsaturated Alkamides; D021122:Protein Subunits; D043882:Receptors, Cannabinoid; C107684:prostaglandin E2 ethanolamide; D007371:Interferon-gamma; D051546:Cyclooxygenase 2; D015232:Dinoprostone; C078814:anandamide