OBJECTIVE: To investigate whether microRNA-615-3p participates in the development and progression of osteoarthritis by regulating chondrogenic differentiation of bone marrow mesenchymal stem cells.
MATERIALS AND METHODS: Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat bone marrow and identified by flow cytometry. After chondrogenic differentiation was induced in BMSCs, expression levels of chondrogenic-specific genes were then detected by quantitate Real-time polymerase chain reaction (qRT-PCR). Expression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Protein expression of SOX9 after overexpression or knockdown of microRNA-615-3p was detected by Western blot, respectively.
RESULTS: MicroRNA-615-3p was down-regulated in the process of chondrogenic differentiation of BMSCs. The mRNA expressions of chondrogenic-specific markers, COL2A1, COL10A1, ACAN and MATN3 were decreased after microRNA-615-3p overexpression in BMSCs. Overexpressed microRNA-615-3p down-regulated protein expression of SOX9. Expression levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-α (IL-α) were increased after overexpression of microRNA-615-3p, while inhibition of microRNA-615-3p expression obtained the opposite result. In addition, overexpression of SOX9 rescued the effect induced by microRNA-615-3p on inflammatory cytokines.
CONCLUSIONS: MicroRNA-615-3p participates in the development and progression of osteoarthritis by increasing the expressions of inflammatory cytokines and inhibiting chondrogenic differentiation of BMSCs.
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