Fig. 9

Mechanism of cangrelor against LPS-induced neuroinflammation in BV-2 cells. A Immunofluorescence staining of GPR17 (red), Iba1 (green), and DAPI (blue) in the BV-2 cells. BV-2 cell viability was examined by CCK8 assay after different concentrations of cangrelor (20, 40, and 80 μM) for 24 h (B) and after LPS (1 µg/ml, 24 h) stimulation with or without cangrelor (C). The levels of TNF-α (D), IL-1β (E), and IL-6 (F) in BV-2 cells treated with LPS (1.0 µg/ml) for 24 h with or without cangrelor pretreatment. The expressions of NF-κB p65, p-CREB, and BDNF were examined by Western blotting in BV-2 cells treated with LPS (1 µg/ml) for 24 h with or without cangrelor pretreatment (G). Quantification of NF-κB p65 (H), p-CREB (I), and BDNF (J) protein levels are expressed as the fold difference relative to the control group. After incubation with GPR17 siRNA or its control siRNA for 48 h, BV-2 cells were administrated with 40 μM cangrelor for 1 h followed by LPS (1.0 µg/ml) for an additional 24 h. The levels of TNF-α (K), IL-1β (L) and IL-6 (M). The expressions of NF-κB p65, p-CREB, and BDNF were examined by western blotting (O). Quantification of NF-κB p65 (P), p-CREB (Q), and BDNF (R) was presented as the ratio (in percentage) of the Veh + Veh group. Data shown are expressed as mean ± SEM; n = 4. *P < 0.05, **P < 0.01 versus Veh + LPS