Exercise promotes osteogenic differentiation by activating the long non-coding RNA H19/microRNA-149 axis

Figure 1 Exercise promotes bone formation in growing mice. A: Hematoxylin-eosin staining of mouse femur; B: Changes in bone morphogenetic protein 2 protein expression in mouse femur by immunohistochemical staining; C: Changes in protein expression of Beclin1 and P62 in mouse femur by immunofluorescence staining; D and E: Reconstruction images and bone morphometric indices of changes in the distal femur in mice by micro-CT scanning. n = 6, ^aP < 0.05, ^bP < 0.01. Con: Control group; Ex: Treadmill exercise group.

Figure 2 Exercise promotes osteogenic differentiation of mouse bone mesenchymal stem cells. A: Alkaline phase staining of mouse bone mesenchymal stem cells (BMSCs) at 7 d of osteogenic differentiation induction; B: Alizarin red staining of mouse BMSCs at 14 d of osteogenic differentiation induction; C and D: Identification of potential miRNAs targets of lncRNA H19 by bioinformatics analysis; E and F: Changes in mRNA levels of lncRNA H19, miR-149, and osteogenic differentiation-associated markers after induction of osteogenic differentiation in mouse primary BMSCs by RT-qPCR; G and H: Changes in protein level of osteogenic differentiation-related marker and autophagy-related marker after induction of osteogenic differentiation in mouse primary BMSCs. n = 3, ^aP < 0.05, ^bP < 0.01. Con: Control group; OI: Osteogenic differentiation induction; Ex: Treadmill exercise group.

Figure 3 Exercise promotes osteogenic differentiation of bone mesenchymal stem cells through upregulation of lncRNA H19/miR-149. A: Alkaline phase staining of bone mesenchymal stem cells (BMSCs) 3 d after osteogenic induction of differentiation; B: Alizarin red staining of BMSCs 3 d after osteogenic induction of differentiation; C: Gene expression changes of lncRNA H19, miR-149 and osteogenic differentiation-related markers in BMSCs after osteogenic differentiation induction and si-lncRNA H19 interference by RT-qPCR; D and E: Protein expression changes of osteogenic differentiation-associated markers and autophagy-associated markers in BMSCs after osteogenic differentiation induction and si-lncRNA H19 interference by WB technique. n = 3, ^aP < 0.05, ^bP < 0.01. Con: Control group; NC: Negative control group; OI: Osteogenic differentiation induction group.

Figure 4  Summary diagram of this study.