title: Adenosine signalling mediates the anti-inflammatory effects of the COX-2 inhibitor nimesulide.
abstract: Extracellular adenosine formation from ATP is controlled by ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase/CD39) and ecto-5'-nucleotidase (e-5NT/CD73); the latter converts AMP to adenosine and inorganic phosphate, representing the rate limiting step controlling the ratio between extracellular ATP and adenosine. Evidence that cellular expression and activity of CD39 and CD73 may be subject to changes under pathophysiological conditions has identified this pathway as an endogenous modulator in several diseases and was shown to be involved in the molecular mechanism of drugs, such as methotrexate, salicylates , interferon-β. We evaluated whether CD73/adenosine/A2A signalling pathway is involved in nimesulide anti-inflammatory effect, in vivo and in vitro. We found that the adenosine A2A agonist, 4-[2-[[6-amino-9-(N-ethyl-β-d-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride (CGS21680, 2mg/kg ip.), inhibited carrageenan-induced rat paw oedema and the effect was reversed by co-administration of the A2A antagonist -(2-[7-amino-2-[2-furyl][1,2,4]triazolo[2,3-a][1,3,5]triazin-5-yl-amino]ethyl)phenol (ZM241385; 3mg/kg i.p.). Nimesulide (5mg/kg i.p.) anti-inflammatory effect was inhibited by pre-treatment with ZM241385 (3mg/kg i.p.) and by local administration of the CD73 inhibitor, adenosine 5'-(α,β-methylene)diphosphate (APCP; 400μg/paw). Furthermore, we found increased activity of 5'-nucleotidase/CD73 in paws and plasma of nimesulide treated rats, 4h following oedema induction. In vitro, the inhibitory effect of nimesulide on nitrite and prostaglandin E2 production by lipopolysaccharide-activated J774 cell line was reversed by ZM241385 and APCP. Furthermore, nimesulide increased CD73 activity in J774 macrophages while it did not inhibit nitrite accumulation by lipopolysaccharide-activated SiRNA CD73 silenced J774 macrophages. Our data demonstrate that the anti-inflammatory effect of nimesulide in part is mediated by CD73-derived adenosine acting on A2A receptors.
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Caiazzo
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Maione
authors.affiliation: Department of Pharmacy, University of Salerno, Fisciano, Salerno, Italy.
authors.forename: Silvana
authors.initials: S
authors.lastname: Morello
authors.affiliation: Department of Health Sciences, Clinical Pharmacology and Oncology Section, University of Florence, Florence, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lapucci
authors.affiliation: Department of Health Sciences, Clinical Pharmacology and Oncology Section, University of Florence, Florence, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Paccosi
authors.affiliation: Department of Molecular Cardiology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Bodo
authors.initials: B
authors.lastname: Steckel
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lavecchia
authors.affiliation: Department of Health Sciences, Clinical Pharmacology and Oncology Section, University of Florence, Florence, Italy.
authors.forename: Astrid
authors.initials: A
authors.lastname: Parenti
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy.
authors.forename: Teresa
authors.initials: T
authors.lastname: Iuvone
authors.affiliation: Department of Molecular Cardiology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schrader
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy.
authors.forename: Armando
authors.initials: A
authors.lastname: Ialenti
authors.affiliation: Department of Pharmacy, University of Naples Federico II, Napoli, Italy. Electronic address: cicala@unina.it.
authors.forename: Carla
authors.initials: C
authors.lastname: Cicala
mesh_terms: D015720:5'-Nucleotidase / Q000037:antagonists & inhibitors / Q000097:blood / Q000378:metabolism; D000241:Adenosine / Q000378:metabolism*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D052246:Cyclooxygenase 2 Inhibitors / Q000008:administration & dosage / Q000494:pharmacology*; D015232:Dinoprostone / Q000097:blood; D004487:Edema / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D008297:Male; D017208:Rats, Wistar; D043705:Receptor, Adenosine A2A / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013449:Sulfonamides / Q000008:administration & dosage / Q000494:pharmacology*
chemical_list: D000893:Anti-Inflammatory Agents; D052246:Cyclooxygenase 2 Inhibitors; D043705:Receptor, Adenosine A2A; D013449:Sulfonamides; D015720:5'-Nucleotidase; D000241:Adenosine; D015232:Dinoprostone; C012655:nimesulide
keywords: -(2-[7-amino-2-[2-furyl][1,2,4]triazolo[2,3-a][1,3,5]triazin-5-yl-amino]ethyl)phenol (ZM241385, PubMed CID: 176407); 4-[2-[[6-amino-9-(N-ethyl-β-d-ribofuranuronamidosyl)-9H-purin-2-yl]amino]ethyl]benzenepropanoic acid hydrochloride (CGS21680, PubMed CID: 10256643); 5′-(α,β-methylene)diphosphate (APCP, PubMed CID: 16219694); Adenosine; Celecoxib (PubMed CID: 2662); Ecto-5′-nucleotidase; Inflammation; J774 macrophages; LPS; Nimesulide; Nimesulide (PubMed CID: 4495)