MicroRNA-590-5p antagonizes the inhibitory effect of high glucose on osteoblast differentiation by suppressing Smad7 in MC3T3-E1 cells

Abstract

Objective: MicroRNA-590-5p (miR-590-5p) has been reported to stimulate osteoblast differentiation; however, its effect in diabetic osteoporosis remains unknown. This study investigated the effect of miR-590-5p on high glucose (HG)-suppressed osteoblast differentiation. Methods: The effect of HG on MC3T3-E1 cell survival was assessed using the MTT assay. The expression levels and activities of osteoblastic proteins were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR), alkaline phosphatase (ALP) assay, and immunoblotting assay. Tumor growth factor-β (TGF-β) signaling in MC3T3-E1 cells was assessed using luciferase assay, qRT-PCR, and immunoblotting. Mineralized nodule formation in MC3T3-E1 cells was examined by using the mineralization assay. Results: When MC3T3-E1 cells were exposed to HG conditions, there was significant downregulation of miR-590-5p and osteoblastic proteins (e.g., collagen I, Runx2, and ALP); in contrast, Smad7 was upregulated. Furthermore, miR-590-5p targeted Smad7 and inhibited its expression. Additionally, overexpression of miR-590-5p significantly promoted osteoblast growth and differentiation by upregulating TGF-β signaling in HG-treated MC3T3-E1 cells. Conclusions: Collectively, the results showed that miR-590-5p was involved in osteogenesis; moreover, miR-590-5p may represent a potential target for the treatment of diabetic osteoporosis.