miR-138-5p inhibits healing of femoral fracture osteogenesis in rats by modulating osteoblast differentiation via SIRT1/FOXO1 axis.

Abstract

Background: MicroRNAs have a crucial part to play in maintaining bone formation, signaling, and repair. This research explored the involvement miR-138-5p in modulating osteoblast differentiation in femoral fractures model.

Methods: The role of mir-138-5p in the healing process of femoral fractures in rats was assessed through micro computed tomography (CT) imaging. After that, qPCR was employed to identify the cellular mRNA expression levels of miR-138-5p, SIRT1, and FoxO1 in either the callus or MC3T3-E1. Next, the protein expression level of Runx2, OPN, OCN and ALP was determined by western blot or ELISA. A dual-luciferase reporter gene assay was implemented to examine the target of miR-138-5p. The quantity of mineralized nodules was measured by means of alizarin red staining.

Results: The miR-138-5p inhibitor promotes the mending of femoral fractures. When it is knocked down, the osteogenic differentiation is promoted, which may be caused by the enhanced activity of ALP and the elevation of the expression of Runx2, OPN and OCN. Meanwhile, an increase in the expression of mir-138-5p impairs the biosynthesis of SIRT1 and FoxO1. When SIRT1 and FoxO1 were downregulated with shRNA, the effect caused by the mir-138-5p inhibitor could be reversed.

Conclusion: Our studies uncovered that the overexpressed miR-138-5p might have an inhibitory role in femoral fractures healing by inactivating SIRT1/FOXO1 axis.