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2022, Vol. 26 ›› Issue (7): 1020-1026Bushen Huoxue capsule regulates osteogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells via Hedgehog signaling pathwayLiang Xuezhen1, Yang Xi2, Li Jiacheng1, Luo Di3, Xu Bo1, Li Gang1, 3
Abstract: BACKGROUND: Steroid-induced osteonecrosis of femoral head, a common, progressive and refractory orthopedics disease, seriously affected the normal life and work of the patients. The imbalance of osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was confirmed to involve in the pathogenesis, prevention, and treatment of steroid-induced osteonecrosis of femoral head. However, the mechanism is unclear. OBJECTIVE: To observe the effect of Bushen Huoxue capsule containing serum on osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells treated with glucocorticoids and Hedgehog signal pathway related factors. METHODS: Bone marrow mesenchymal stem cells were isolated from SD rats and cultured, and intervened with different doses of Bushen Huoxue capsule containing serum and glucocorticoids. CCK-8 assay was used to observe the proliferation of cells. The osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells in different groups were detected by alizarin red staining and oil red “O” staining. The changes of protein expression of osteogenic factors and adipogenic factors and Hedgehog signal pathway related factors of bone marrow mesenchymal stem cells in different groups were analyzed by western blot assay. RESULTS AND CONCLUSION: (1) The Bushen Huoxue capsule containing serum not only could promote the proliferation of normal rat bone marrow mesenchymal stem cells, but also could obviously increase the proliferation activity of bone marrow mesenchymal stem cells treated by the glucocorticoids. (2) After 3 weeks of bone induction, osteogenic induction abilities in the low dose group, middle dose group and high dose group of Bushen Huoxue capsule containing serum were significantly higher than that of the blank control group, especially the middle dose group. (3) After 2 weeks of lipid induction, the positive staining areas revealed by oil red “O” staining in the low dose group, middle dose group and high dose group of Bushen Huoxue capsule containing serum were lower than that of the blank control group, especially the middle dose group. (4) After 3 days of intervention in each group, western blot assay showed that Bushen Huoxue capsule could enhance the protein expression of osteogenic factors (Runx2, Col1a1) and Hedgehog signal pathway related factors (Shh, Gli1) in bone marrow mesenchymal stem cells, and weaken the protein expression of adipogenic factors (PPARγ, C/EBPα) in bone marrow mesenchymal stem cells. (5) The results indicate that the Bushen Huoxue capsule containing serum could promote the osteogenic differentiation of bone marrow mesenchymal stem cells and inhibit the adipocyte differentiation of bone marrow mesenchymal stem cells through the Hedgehog signaling pathway. Key words:stem cells, bone marrow mesenchymal stem cells, steroid-induced osteonecrosis of the femoral head, Bushen Huoxue capsule, Hedgehog signal pathway, osteogenic differentiation, adipogenic differentiation |