Abstract
Our previous studies have shown adenosine A2A R activation markedly promotes the expression of cystatin F (CF) and exacerbates the white matter lesions induced by hypoxic brain injuries. Thus, we hypothesized that CF was probably involved in neuroinflammation of activated microglia induced by A2A R activation. We transfected the BV2 cells with a CF shRNA vector and examined the production of pro-inflammatory cytokines in hypoxic-BV2 cells in which A2A R was activated or inactivated to confirm this hypothesis. Additionally, we also investigated the probable signaling pathways involved in modulation of A2A R activation on CF expression in hypoxia-activated BV2 cells. Activation of A2A R promoted CF expression, which was significantly increased after the low glucose and hypoxia treatments in BV2 cells. CF gene knockdown markedly inhibited the increase in the expression of pro-inflammatory cytokines induced by A2A R activation in hypoxic-BV2 cells. Furthermore, the increased expression of the CF induced by A2A R activation was remarkably inhibited in hypoxic-BV2 cells administrated with the PKA inhibitor H-89 and the PKC inhibitor staurosporine. Hence, these results indicate that hypoxia BV2 cells highly express CF, which is involved in A2A R activation-mediated neuroinflammation via the PKA/CREB and PKC/CREB or ERK1/2 signaling pathways.
Highlights
Adenosine, which activates the A2A receptor (A2A R), has been reported to play an important role in regulating the inflammatory response in various types of brain injuries[1,2,3]
We showed that A2A R activation by CGS21680 promoted cystatin F (CF) expression, which was significantly increased after the low glucose and hypoxia treatments
We inhibited CF expression in BV2 cells by transfecting the cells with a CF short hairpin RNA (shRNA) viral vector and showed that the increased expression of CF may be involved in the effects of A2A R activation on the increased production of pro-inflammatory cytokines in hypoxic BV2 cells in response to the low glucose and hypoxia treatments
Summary
Adenosine, which activates the A2A receptor (A2A R), has been reported to play an important role in regulating the inflammatory response in various types of brain injuries[1,2,3]. CF expression was substantially up-regulated in regions of white matter rarefaction that occurred in various demyelinating diseases of the central nervous system[7,8] Consistent with these findings, our previous study showed that CF was expressed in activated microglial cells in white matter lesions induced by chronic cerebral hypoperfusion. A2A R inactivation substantially increased CF production in activated microglial cells; the production of inflammatory cytokines was significantly increased in white matter lesions after chronic cerebral hypoperfusion[9]. These results suggested that CF expressed in activated www.nature.com/scientificreports/.
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