长链非编码RNA AK146527通过靶向miR-15/16调控骨形成的分子机制OA

Objective:To investigate the regulatory role of AK146527 in osteogenic differentiation and bone formation,explore the molecular mechanism by which AK146527 promotes bone formation through targeting miR-15/16,and assess the in situ osteogenic effect of the functional region of AK146527.Methods:RT-PCR was performed to detect the expression levels of AK146527 in mouse femoral tissues,and the correlation between AK146527 expression and bone formation markers was analyzed.MC3T3-E1 pre-osteoblasts were transfected with AK146527 overexpression plasmids or siRNA to evaluate the effect of AK146527 on osteoblast differentiation.The activity of key transcription factors involved in osteogenic differentiation was preliminarily screened in MC3T3-E1 cells after AK146527 transfection to explore the specific signaling pathways through which AK146527 regulates osteogenic differentiation.MC3T3-E1 cells were transfected with AK146527 overexpression plasmids or siRNA,and the levels of miR-15/16 family members were measured to investigate the targeted regulation of the miR-15/16 family by AK146527.MC3T3-E1 cells and C57BL/6 mice were transfected with overexpression plasmids targeting different regions of AK146527,and osteogenic differentiation levels in cells and bone formation levels in mice were assessed to confirm that the binding sequence of AK146527 with miR-15/16 regulates osteoblast differentiation and bone formation.An ovariectomized(OP)mouse model was established,and overexpression plasmids containing the AK146527 binding region were locally transfected into the subcutis of the mouse skull to evaluate the therapeutic effect of the AK146527 binding region on OP.Finally,the in situ osteogenic effect of the AK146527 binding region was validated using organoid technology.Results:The expression level of AK146527 was correlated with bone formation.AK146527 promoted bone formation.The regulatory effect of AK146527 on osteogenic differentiation may involve the Wnt signaling pathway.AK146527 specifically bound to and inhibited the miR-15/16 family through its binding region.AK146527 regulated osteogenic differentiation via the miR-15/16 binding site.The binding region of AK146527 exhibited in situ osteogenic effects and demonstrated promising potential for application in bone defect repair.Conclusion:AK146527 is highly correlated with bone formation and promotes osteogenic differentiation and bone formation by targeting miR-15/16.Its binding region with miR-15/16 can restore bone formation in ovariectomized osteoporotic mice.Additionally,the functional region of AK146527 exhibits in situ osteogenic and bone repair effects after implantation into organoids in a mouse skull defect model.This study provides a novel theoretical and experimental foundation for understanding the mechanisms of bone formation and offers a new strategy to enhance the regulatory efficiency of bone formation-related LncRNAs.