Interferon-gamma inhibits adenosine A2A receptor function in hepatic stellate cells by STAT1-mediated repression of adenylyl cyclase.

BACKGROUND AND PURPOSE: Adenosine, an endogenous purine nucleoside, is a potent regulator of the inflammatory response and stimulus for fibrosis. We have previously demonstrated that adenosine, acting at the A2A receptor, plays a central role in hepatic fibrosis via direct promotion of collagen production by hepatic stellate cells. As we have previously demonstrated that macrophage A2A receptor function is regulated by interferon-gamma (IFNγ), a noted antifibrotic but pro-inflammatory cytokine, we examined its effect on A2AR-stimulated collagen production in the human hepatic stellate cell line LX-2. EXPERIMENTAL APPROACH: Collagen expression was determined by western blotting and realtime reverse transcription polymerase chain reaction (RT-PCR). Receptor desensitization was assessed by western blotting for membrane associated GRK2. Receptor signaling was determined by western blotting for phosphorylated extracellular signal-related protein kinase (ERK) protein and immunoassay for intracellular cyclic AMP (cAMP). siRNA was used to knock down expression of adenylyl cyclase and signal transducer and activator of transcription (STAT). Adenylyl cyclase expression was assessed by realtime RT-PCR, and STAT expression was assessed by western blotting. KEY RESULTS: IFNγ diminishes A2A receptor-mediated collagen production at both protein and mRNA levels. IFNγ alters signal transduction at A2A receptors by a STAT1 mediated mechanism involving the suppression of adenylyl cyclase expression. CONCLUSIONS AND IMPLICATIONS: IFNγ inhibits the function of the adenosine A2A receptor in hepatic stellate cells by downregulating the expression of adenylyl cyclase. This finding explains, at least in part, the protective effect of IFNγ in hepatic fibrosis.