| Objective To investigate the effect of SDF-1/CXCR4 pathway on myogenesis of bone marrow mesenchymal stem cells (BMSCs) and osteogenic differentiation of myogenic stem cells (MDSCs) under the intervention of invigorating kidney and invigorating spleen. Methods Normal serum, conditional serum (normal serum plus corresponding osteogenic and muscle-forming induction fluid), and kidney-Yin, kidney-Yang, invigorating spleen, and invigorating kidney-spleen serum were used in BMSCs muscle formation and MDSCs bone differentiation cultures for 15 days. The expression of Desmin was determined with immunofluorescence. The average fluorescence intensity of cells was analyzed with Image j. Mineralization nodules were observed with Alizarin red staining. The expressions of myosin, BMP2, SDF-1 and CXCR4 were detected with enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of SDF-1 and CXCR4 were detected with real-time quantitative PCR. Results Compared to those in normal group, myosin protein level decreased (P<0.05), Desmin expression increased (P<0.01), and SDF-1, CXCR4 protein and mRNA expressions increased in induction group during myoblast differentiation of BMSCs. In the osteogenic differentiation of MDSCs, small area of mineralized nodules appeared in the induction group. The expressions of BMP2 protein, SDF-1 and CXCR4 protein and mRNA increased (P<0.05). Compared to those in the induction group, Desmin expression, myosin protein level, and SDF-1 and CXCR4 protein and mRNA expression levels increased significantly in the treatment group during myoblast differentiation of BMSCs (P<0.01). During the osteogenic differentiation of MDSCs, the number and area of mineralized nodules increased in the treated group. The protein level of BMP2, protein and mRNA expressions of SDF-1 and CXCR4 increased (P<0.05). Conclusion Kidney-invigorating spleen method enhances the protein and mRNA expressions of SDF-1/CXCR4 pathway, thus promoting BMSCs muscle formation and MDSCs bone differentiation. Kidney-invigorating spleen method has the most obvious promoting effect. |