Cystatin F involvement in adenosine A2A receptor-mediated neuroinflammation in BV2 microglial cells

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.

Adenosine A2A Receptors in Bone Marrow-Derived Cells Attenuate Cognitive Impairment in Mice After Chronic Hypoperfusion White Matter Injury

“…CF, a potent endogenous cysteine protease inhibitor, was substantially up-regulated in regions of white matter rarefaction that occurred in various demyelinating diseases of the CNS [32,33]. In this study, we found that BMDC A 2A Rs suppressed CF-mediated microglia activated and non-BMDC A 2A Rs promoted CF-mediated microglia activated, which was consistent with our previous research [16]. The immunohistochemistry suggested BBB alterations was not the key point for cognitive function recovery with adenosine A 2A Rs.…”

“…Our previous research confirmed that BMDC A 2A Rs can inhibit inflammation [15], while non-BMDC A 2A Rs activate microglia and promote inflammatory responses [16]. Different sources of adenosine A 2A R play contradictory roles.…”

Adenosine A2A Receptor Suppressed Astrocyte-Mediated Inflammation Through the Inhibition of STAT3/YKL-40 Axis in Mice With Chronic Cerebral Hypoperfusion-induced White Matter Lesions

“…Finally, astrocytes with a purity of more than 95% were obtained after 3 days of culture. After CGS21680 (100 nM), SCH58261 (50 nM) ( 18 ), Colivelin (10 nM) ( 19 ), and Stattic (50 nM) ( 20 ) had been added separately, the primary astrocytes were cultured for 72 h under mild oxygen-glucose deprivation (mOGD) conditions; that is, in low-glucose Dulbecco’s modified Eagle’s medium/F12 medium (1 g/L) in a hypoxia incubator (3% O 2 , 5% CO 2 , 92% N 2 ). Subsequently the expression levels of the target factors were determined.…”

Necrosulfonamide exerts neuroprotective effect by inhibiting necroptosis, neuroinflammation, and α-synuclein oligomerization in a subacute MPTP mouse model of Parkinson’s disease

“…6 d–g; iNOS, F 3, 20 = 16.22, p < 0.01; IL-1β, F 3, 20 = 19.25, p < 0.01; TNF-α, F 3, 20 = 19.35, p < 0.01). Moreover, NSA inhibited the expression of cystatin F (CST7), a key modulator of inflammatory responses in microglia 39 , 40 (Fig. 6 d,h; F 3, 16 = 21.06, p < 0.01).…”