A2A receptor-induced overexpression of pannexin-1 channels indirectly mediates adenosine fibrogenic actions by favouring ATP release from human subcutaneous fibroblasts

Aims: Disorganization of the subcutaneous tissue due to inflammation and fibrosis is a common feature in patients with myofascial pain. Dermal accumulation of adenosine favours collagen production by human subcutaneous fibroblasts (HSCF) via A(2A) receptors (A(2A)R) activation. Adenosine mimics the fibrogenic effect of inflammatory mediators (e.g. histamine, bradykinin), which promote ATP release from HSCF via plasma-membrane-bound pannexin-1 (Panx1) and/or connexin-43 (Cx43) channels, but this mechanism has never been implicated in A(2A)R actions. Materials and methods: A(2A)R-mediated effects on Panx1 and Cx43 protein amounts were evaluated in primary cultures of HSCF by confocal microscopy and Western blot analysis. Functional repercussions in collagen production, intracellular [Ca2+]i oscillations and ATP release were also evaluated. Key findings: NECA and CGS21680, two enzymatically-stable A(2A)R agonists, increased Panx1, but reduced Cx43, protein density in HSCF. This effect was accompanied by increases in ATP release and collagen III production by HSCF. The involvement of the A(2A)R was confirmed by blockage with the selective A(2A)R antagonist, SCH442416. Inhibition of Panx1 channels by probenecid and the Panx1 mimetic inhibitory peptide, 10Panx, also decreased ATP release and collagen production by HSCF under similar conditions. Superfluous ATP release by HSCF exposed to A(2A)R agonists overexpressing Panx1 channels contributes to keeping high [Ca2+]i levels when the cells were exposed to histamine. Significance: Adenosine A(2A)R-induced Panx1 overexpression was shown here for the first time in HSCF; this feature indirectly implicates ATP release in the fibrogenic vicious cycle operated by adenosine accumulating in subcutaneous tissue fibrosis and myofascial pain associated to dermal inflammation.

Automated summary

This study demonstrates for the first time the overexpression of Panx1 in HSCF induced by A(2A)R, indirectly implicating ATP release in the fibrogenic vicious cycle operated by adenosine accumulating in subcutaneous tissue fibrosis and myofascial pain associated with dermal inflammation.