Expression and functional role of adenosine receptors in regulating inflammatory responses in human synoviocytes |
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Authors: | K Varani F Vincenzi A Tosi M Targa FF Masieri A Ongaro M De Mattei L Massari PA Borea |
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Affiliation: | 1.Department of Clinical and Experimental Medicine, Orthopaedic Clinic, University of Ferrara, Ferrara, Italy;2.Department of Morphology and Hystology, Orthopaedic Clinic, University of Ferrara, Ferrara, Italy;3.Department of Biomedical Sciences and Advanced Therapies, Orthopaedic Clinic, University of Ferrara, Ferrara, Italy |
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Abstract: | Background and purpose:Adenosine is an endogenous modulator, interacting with four G-protein coupled receptors (A1, A2A, A2B and A3) and acts as a potent inhibitor of inflammatory processes in several tissues. So far, the functional effects modulated by adenosine receptors on human synoviocytes have not been investigated in detail. We evaluated mRNA, the protein levels, the functional role of adenosine receptors and their pharmacological modulation in human synoviocytes.Experimental approach:mRNA, Western blotting, saturation and competition binding experiments, cyclic AMP, p38 mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB activation, tumour necrosis factor α (TNF-α) and interleukin-8 (IL-8) release were assessed in human synoviocytes isolated from patients with osteoarthritis.Key results:mRNA and protein for A1, A2A, A2B and A3 adenosine receptors are expressed in human synoviocytes. Standard adenosine agonists and antagonists showed affinity values in the nanomolar range and were coupled to stimulation or inhibition of adenylyl cyclase. Activation of A2A and A3 adenosine receptors inhibited p38 MAPK and NF-κB pathways, an effect abolished by selective adenosine antagonists. A2A and A3 receptor agonists decreased TNF-α and IL-8 production. The phosphoinositide 3-kinase or Gs pathways were involved in the functional responses of A3 or A2A adenosine receptors. Synoviocyte A1 and A2B adenosine receptors were not implicated in the inflammatory process whereas stimulation of A2A and A3 adenosine receptors was closely associated with a down-regulation of the inflammatory status.Conclusions and implications:These results indicate that A2A and A3 adenosine receptors may represent a potential target in therapeutic modulation of joint inflammation. |
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Keywords: | adenosine receptors, human synoviocytes, mRNA, Western blotting, receptor binding, cAMP, MAPK p38, NF-κ B, TNF-α , IL-8 |
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