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. 2016 Dec 9;6:38622. doi: 10.1038/srep38622

Figure 2. Oral administration of CAPE attenuates MSU crystals-induced NLRP3 inflammasome activation in a mouse air pouch model.

Figure 2

(A–E) Air pouches were formed on the dorsa of C57BL/6 mice by injecting air twice. The mice were orally administered CAPE (30 mg/kg) or vehicle (Veh, 0.02% DMSO in water). After 1 hr, MSU crystals (3 mg/ml in PBS/mouse) or PBS alone were injected into the air pouches. After 6 hr, the pouch exudates were harvested and the supernatants were analyzed by (A) immunoblotting for caspase-1(p10) and IL-1β, (B) caspase-1 enzyme activity assay, and ELISAs for (C) IL-1β, and (D) IL-18. (E) Bone marrow-derived immortalized macrophages that had been transfected with the iGLuc luciferase reporter plasmid were injected into the air pouches. After 3 hr, the mice were orally administered CAPE (30 mg/kg) or vehicle. After 1 hr, MSU crystals (3 mg/ml in PBS/mouse) or PBS alone were injected into the air pouches. After 6 hr, luminescence derived from iGLuc-luciferase expression was assessed by in vivo imaging analysis using an Xtreme system (Bruker). (F) The air pouch tissue was fixed for histological examination using H&E staining. The purple dots represent infiltrated neutrophils. (G) Myeloperoxidase (MPO) activity, which reflects neutrophil recruitment, was assessed in the air pouch exudates. The values in the bar graphs represent the means ± SEM (n = 3–6 mice). #Significantly different from vehicle alone, p < 0.05. *Significantly different from MSU alone, p < 0.05. Veh, vehicle.