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URL: https://dx.doi.org/DOI:10.17504/protocols.io.vzhe736

Authors: Juan Wu

Summary: In the current study, we successfully established an acute inflammation model and a chronic inflammation model involving adipose stem cells. We used high-throughput miRNA microarray analysis to identify miRNAs that were significantly (pwhich was directly targeted by miR-223-3P. STAT3 directly targeted TLR4 and TLR2, promoting the production of the pro-inflammatory cytokine, IL-6, and formed a positive feedback loop to regulate IL-6 levels. Similarly, TNF-a significantly(p1.Establish an acute inflammation model of adipose stem cells2.Establish a chronic inflammation model of adipose stem cells3. miRNA microarray and data analysis4.Target gene prediction, gene ontology (GO) enrichment, and pathway analysis5.Prediction of target gene binding sites6.Dual luciferase assay7.LPS induces STAT3mRNA and protein levels in adipose stem cells, and promotes STAT3 protein phosphorylation, while miR-223-3p regulates STAT3 mRNA8.miR-223-3p directly regulates TLR2 and TLR4, and the secretion of inflammatory cytokines IL-6, TNF-a, and IL-1b9. STAT3 promotes phosphorylation of STAT3 protein, and regulates TLR4 and TLR2 expression10.Treatment of adipose stem cells with IL-6, TNF-a, siIL-6, and siTNF-a affects the miR-223-3p, STAT3, and pSTAT3 levels11.miR-2909 depends on NF-kB to target KLF4 to regulate IL-6, IL-1b, and TNF-a levels

Associated Publications: Wu J, Niu P, Zhao Y, Cheng Y, Chen W, Lin L, Lu J, Cheng X, Xu Z (2019) Impact of miR-223-3p and miR-2909 on inflammatory factors IL-6, IL-1ß, and TNF-α, and the TLR4/TLR2/NF-κB/STAT3 signaling pathway induced by lipopolysaccharide in human adipose stem cells. PLoS ONE 14(2): e0212063. doi: 10.1371/journal.pone.0212063

Affiliations: Wuhan Hubei China

External URL: https://doi.org/10.1371/journal.pone.0212063

Version: 2

Publication Date: 2018

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