Cobalt Chloride Enhances the Anti-Inflammatory Potency of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells through the ERK-HIF-1α-MicroRNA-146a-Mediated Signaling Pathway

Figure 3

Cobalt chloride- (CoCl₂-) induced expression of miR-146a in immunomodulation of hUCB-MSCs. (a) hUCB-MSCs (S1–2) and coculture of hPBMCs with hUCB-MSCs (hPBMC + hMSC, S3–4) were collected for microRNA (miR) microarray analysis. Normalized log2 miRNA expression for miRNAs that showed significant differences () between groups is listed according to the color scale. (b) hUCB-MSCs were treated with 100 μM CoCl₂ and then harvested at the indicated times. The expression level of miR-146a was determined by RT-qPCR. (c) hUCB-MSCs were transfected with scrambled miR-control, miR-146a mimics, or inhibitors. The expression level of miR-146a was determined by RT-qPCR. (d–f) hUCB-MSCs were transfected with scrambled miR-control, miR-146a mimics, or inhibitors and exposed to 100 μM CoCl₂ for 72 h, and then allogeneic hPBMCs were cocultured with hUCB-MSCs. PGE₂, TNF-α, and IFN-γ levels in the culture supernatants were measured by ELISA. Data represent the , ; . P: hPBMCs; P: allogeneic hPBMCs as stimulator; H: phytohemagglutinin; MSC: naïve hUCB-MSCs; CoMSC: CoCl₂-pretreated hUCB-MSCs.

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