fig5

Figure 5. UCMSC-NVs attenuate macrophage infiltration in ALI mouse lungs via modulation of the NF-κB/NLRP3 signaling pathway. (A) Protein expression levels of IκBα, p-IκBα, total p65, p-p65, NLRP3, caspase-1, IL-18 and IL-1β were assessed by Western blotting; (B) Statistical analysis of histograms against the WB images in panel A. n = 3 per group; (C and D) Immunofluorescence analysis of M1 macrophage content in lung tissue (n = 6 per group) and M2 macrophage content in lung tissue. n = 6 per group. Scale bar = 100 μm; (E) Flow cytometry analysis of the proportions of neutrophils (CD11b+ Ly6G+) and macrophages (CD11b+ F4/80+) in BALF. n = 3 per group. Data represent means ± SD. Statistical analysis was performed by one-way ANOVA. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. UCMSC-NVs: Nanovesicles originating from human umbilical cord mesenchymal stem cells; ALI: acute lung injury; NF-κB: nuclear factor κB; NLRP3: NOD-like receptor protein 3; p-IκBα: phosphorylated IκBα; p-p65: phosphorylated p65; IL-18: interleukin-18; IL-1β: interleukin-1β; WB: Western blot; BALF: bronchoalveolar lavage fluid; SD: standard deviation; ANOVA: analysis of variance; LPS: lipopolysaccharide; DAPI: 4′,6-diamidino-2′-phenylindole.