Bone Mesenchymal Stem Cell-Derived Extracellular Vesicles Containing Long Noncoding RNA NEAT1 Relieve Osteoarthritis

Figure 5

NEAT1 binds to miR-122-5p, thus upregulating Sesn2 expression and activating the Nrf2 pathway. (a) Differential expression of mRNAs in OA samples in OA-related GSE16464 microarray. (b) Venn diagram of the predicted downstream mRNAs of miR-122-5p by the starBase, TargetScan, and miRDB databases and the downregulated mRNAs in OA samples in the OA-related GSE16464 microarray. (c) Expression of Sesn2 in OA samples in the GSE16464 microarray. (d) Binding sites between Sesn2 and miR-122-5p predicted by TargetScan and their binding in HEK-293T cells confirmed by dual-luciferase reporter assay. (e) Interaction among NEAT1, miR-122-5p, and Sesn2 detected by dual-luciferase reporter assay. (f) Sesn2 expression in cartilage tissues of OA patients () and normal tissues () detected by RT-qPCR (left) as well as the correlation analysis of NEAT1 expression and Sesn2 expression in cartilage tissues of OA patients () by Pearson’s correlation coefficient (right). (g) Sesn2 protein expression in chondrocytes transfected with miR-122-5p mimic or miR-122-5p inhibitor determined by Western blot analysis. (h) Protein expression of the Nrf2 pathway-related factors in chondrocytes transfected with si-Nrf2 or si-NEAT1 determined by Western blot analysis. (i) mRNA expression of Sesn2, Srx1, and Trx1 in chondrocytes treated with oe-NEAT1 or combined with miR-122-5p mimic detected by RT-qPCR. (j) Protein expression of Sesn2, Nrf2, Srx1, and Trx1 in chondrocytes treated with oe-NEAT1 or combined with miR-122-5p mimic detected by Western blot analysis. . Measurement data were expressed as . Data between groups were compared using independent sample test. Correlation was analyzed by Pearson’s correlation analysis. Data among groups were compared by one-way ANOVA followed by Tukey’s post hoc test. The cell experiment was run in triplicate independently.

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