Adenosine kinase inhibitor attenuates the expression of inducible nitric oxide synthase in glial cells

• Published in: Neuropharmacology 2005, Vol.48 (1), p.151-160
• Main Authors: Lee, Jin-Koo, Won, Je-Seong, Singh, Avtar K., Singh, Inderjit
• Format: Article
• Language: English
• Subjects: Adenosine; Adenosine - metabolism; Adenosine Kinase - antagonists & inhibitors; Adenosine kinase inhibitor; Adenosine receptor; Analysis of Variance; Animals; Blotting, Northern - methods; Blotting, Western - methods; C6 glioma cells; Cell Line, Tumor; Cyclic AMP - metabolism; Dose-Response Relationship, Drug; Electrophoretic Mobility Shift Assay - methods; Enzyme-Linked Immunosorbent Assay - methods; Gene Expression Regulation, Enzymologic - drug effects; Glioma; Inducible nitric oxide synthase; Interferon-gamma - pharmacology; L-Lactate Dehydrogenase - metabolism; Leupeptins - metabolism; Lipopolysaccharides - pharmacology; Neuroglia - drug effects; Neuroglia - metabolism; NF-kappaB-Inducing Kinase; Nitric Oxide - metabolism; Nitric Oxide Synthase - genetics; Nitric Oxide Synthase - metabolism; Nitric Oxide Synthase Type II; Proline - analogs & derivatives; Proline - pharmacology; Protein Kinase Inhibitors - pharmacology; Protein Serine-Threonine Kinases - antagonists & inhibitors; Protein Serine-Threonine Kinases - metabolism; Rats; Thiocarbamates - pharmacology; Transfection - methods; Tumor Necrosis Factor-alpha - metabolism
• ISSN: 0028-3908; 1873-7064
• DOI: 10.1016/j.neuropharm.2004.09.006

The present study demonstrates the anti-inflammatory effect of adenosine kinase inhibitor (ADKI) in glial cells. Treatment of glial cells with IC51, an ADKI, stimulated the extracellular adenosine release and reduced the LPS/IFNγ-mediated production of NO, and induction of iNOS and TNF-α gene expression. The recovery of IC51-mediated inhibition of iNOS expression by adenosine transport inhibitor, S-(4-nitrobenzyl)-6-thioinosine (NBTI), and the inhibition of LPS/IFNγ-induced iNOS gene expression by exogenous adenosine indicate a role for adenosine release in IC51-mediated iNOS expression. The rescue of IC51-mediated inhibition of iNOS expression by adenosine receptor antagonist for A 2A, 8-(3-chlorostyryl)caffeine (CSC) and alloxazine for A 2B, further supports a role for interaction of adenosine and its receptors in anti-inflammatory activity. The IC51-mediated induction of cAMP levels, downstream target of A 2A and A 2B, and inhibition of LPS/IFNγ-induced expression of iNOS by forskolin, a cAMP activator, document a role for cAMP mediated pathway in anti-inflammatory activity of IC51. Taken together, these studies document that IC51-mediated inhibition of iNOS expression is through activation of adenosine receptors, which activates A 2A and A 2B resulting in increased cAMP levels following LPS/IFNγ stimulation. Moreover, the lack of effect of IC51 or adenosine on NFκB DNA binding activity and its transactivity indicates that the inhibition of iNOS expression mediated by IC51 may be through an NFκB independent pathway.