Fig. 5

The modulation of miR-653-5p on IL-6/JAK/STAT3 signaling pathway. a GSEA analysis demonstrating JAK/STAT signaling pathway enriched in OA. b KEGG analysis demonstrating JAK/STAT pathway enriched in OA. c Cultured primary human OA chondrocytes were transfected with miR-653-5p mimics, miR-653-5p inhibitor and their negative controls for 72 h and then the levels of IL-6, JAK1, p-JAK1, STAT3, p-STAT3, p16 INK4a , MMP13 and Col2A1 were measured by western blotting. Quantitative analysis was shown on the right, n  = 3 independent biological replicates per group. * * * p < 0.001 by one-way ANOVA test followed by Tukey’s post hoc test. d The rescue experiment was established in cultured primary human OA chondrocytes to validate the relationship between miR-653-5p and IL-6. Inhibition of MMP13, p16 INK4a and IL-6 expression levels by miR-653-5p mimics was rescued by restoration of IL-6 expression. In comparison, inhibition of Col2A1 expression by IL-6 overexpression was rescued by miR-653-5p mimics. Quantitative analysis was shown on the right, n = 3 independent biological replicates per group. * * p < 0.01, * * * p < 0.001 by one-way ANOVA test followed by Tukey’s post hoc test. miRNA, microRNA; IL-6, interleukin 6; Col, collagen; MMP13, matrix metallopeptidase 13; JAK, janus kinase; OA, osteoarthritis; NC, normal control