Fig. 6: CA-SBEβCD NPs-mediated inhibition of proinflammatory cytokines is involved in the regulation of CEBPβ in the brains of APP/PS1 mice.
CA-SBEβCD NPs (loaded with 10 or 30 mg/kg CA) or SBEβCD NPs (vehicle control) were administered to 4-month-old APP/PS1 Tg mice and age-matched WT C57BL/6 mice for 5 months. A Western blot and quantitative analysis showing the protein levels of pro- and mature IL-1β, IL-6 and TNFα in the mouse brain. Real-time PCR analysis showing the mRNA levels of TNFα (B) and IL-6 (C). D Western blot analysis of NFκB p65 expression in the cytosol and nucleus in protein lysates from Tg and WT mice brains. The protein expression levels of COX-2 and NLRP3 were analysed. Representative blots showing the protein levels of cytosolic and nuclear CEBPβ. The mRNA levels of NFκB (E) and CEBPβ (F). G Representative images of immunofluorescence (IF) staining for CEBPβ (red) and GFAP (green) in the hippocampi of WT and Tg mice. The nuclei were labelled with DAPI (blue). The white arrows indicate colocalization of CEBPβ and GFAP. Scale bars: 60 µm. The enlarged images in the right panels show GFAP- and CEBPβ-positive cells. Scale bars: 10 µm. H IF labelling of CEBPβ and Iba1 in the hippocampus of Tg mice. Scale bars: 60 µm. The white arrowheads indicate the colocalizations of CEBPβ and Iba1. Right panels: enlarged images of the indicated cells. Scale bars: 10 µm. I The interaction of CEBPβ with NFκB was assessed by Co-IP. ChIP was performed to assess the NFκB-mediated transcription of IL-6 (J) and TNFα (K). *p < 0.05, **p < 0.01 and ***p < 0.001 vs vehicle-treated mice; ###p < 0.001 vs the WT mice by the two-way ANOVA with post hoc Fisher’s LSD tests. Values represent the mean ± SD. n = 6 in each group. Blots were repeated at least three times for every condition.
