Fig. 5

Knockdown and inhibition of hippocampal GPR17 inhibits neuroinflammation induced by LPS in mice. Representative images of immunofluorescent staining of Iba1 (green) and DAPI (blue) in the hippocampal DG of lentivirus (A) and cangrelor (B) pretreatment mice. Scale bars = 100 μm. The number of Iba1 antibody-stained microglia, in the DG of LV-GPR17–shRNA (C) and cangrelor (D) pretreatment mice. Iba-1 and CD68 protein levels were detected by Western blotting in the hippocampus of lentivirus (E) and cangrelor (F) pretreatment mice. Protein band intensity was normalized to β-actin. G–J Quantification of Iba-1 and CD68 protein levels is expressed as the fold difference relative to the control group. Representative images of GFAP (red) immunofluorescent staining in the hippocampal DG of the LV-GPR17–shRNA (K) and cangrelor (L) pretreatment groups. Scale bars = 100 μm. M, N Number of GFAP antibody-stained astrocytes was normalized, as the ratio (in percentage) of the Veh + Veh is shown. GFAP protein levels were detected by Western blotting in the mice of pretreatment with LV-GPR17–shRNA (O) and cangrelor (P). Protein band intensity was normalized to β-actin. Q, R Quantification of GFAP protein levels is expressed as the fold difference relative to the control group. S–U Pro-inflammatory cytokine TNF-α, IL-1β, and IL-6 in the brain were measured by ELISA in the mice of pretreatment with LV-GPR17–shRNA. V–X Pro-inflammatory cytokine TNF-α, IL-1β, and IL-6 in the brain were measured by ELISA in the mice of pretreatment with cangrelor. Data shown are expressed as mean ± SEM; n = 4 mice/group. *P < 0.05, **P < 0.01 versus Veh + LPS