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. 2019 Aug 22;4(16):e125437. doi: 10.1172/jci.insight.125437

Figure 1. ANGPTL4 derived from MSCs is essential for inhibiting the inflammatory activation of macrophages.

Figure 1

(A) Both ANGPTL4 mRNA and protein were notably increased in human mesenchymal stem cells (hMSCs) when cocultured with either unstimulated human primary macrophages (hMFs) or LPS-stimulated hMFs. n = 4. (B) siRNA control–transfected (siCon-transfected) hMSCs reduced the mRNA levels of inflammatory markers Cxcl9, Cxcl10, Cxcl11, Ccr7, Il6, and Ccl2 in LPS-stimulated hMFs after coculture for 24 hours, while siRNA ANGPTL4–transfected (siANGPTL4-transfected) hMSCs did not. n = 4. (C) Inflammatory markers in LPS-stimulated hMFs were significantly reduced by treatment with recombinant ANGPTL4 protein. n = 4. (D) Differentiated THP-1 macrophages showed significant reductions in proinflammatory CXCL10 mRNA (n = 4) and protein induction (n = 8) by LPS stimulation by coculture with hMSCs. (E) LPS-induced degradation of IκBα protein in THP-1 macrophages was blunted by coculture with hMSCs. Western blots are representative of n > 3 repeats. Intensity quantification is representative of mean ± SEM. (F–H) ANGPTL4 mRNA (n = 4), cellular protein (n = 4), and released protein (n = 4) were assayed in hMSCs with or without coculture with LPS-stimulated THP-1 macrophages. (I) ANGPTL4 protein–treated THP-1 macrophages showed significant reductions in inflammatory genes in a dose-dependent manner. n = 4. Data are represented as mean ± SEM. #P < 0.05; ##P < 0.01; ###P < 0.001 (by Student’s t test or 1-way ANOVA with Bonferroni’s multiple-comparisons test).