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目的:应用体外培养骨髓间充质细胞,在细胞和分子水平观察补骨合剂对细胞分化的影响,探讨补骨合剂治疗骨质疏松症的机制。方法:分离SD大鼠骨髓间充质细胞,通过形态学、碱性磷酸酶、细胞内骨钙素、转化生长因子β1基因表达等指标,对重组人骨形态发生蛋白2(rhBMP2)诱导下的骨髓间充质细胞向成骨细胞分化的程度进行探讨。结果:一定浓度的补骨合剂可促进分化中的骨髓间充质细胞分泌碱性磷酸酶和骨钙素,增强rhBMP2的活性,促进分化中的骨髓间充质细胞表达转化生长因子β1,补骨合剂的这种促进骨髓间充质细胞分化作用与密钙息基本相同。结论:补骨合剂不仅可以增强rhBMP2的活性,促进分化中的骨髓间充质细胞表达转化生长因子β,大量分泌Ⅰ型胶原,以利于钙盐沉积;还可以促进rhBMP2诱导的骨髓间充质细胞分泌碱性磷酸酶和骨钙素,促进骨髓间充质细胞向成骨细胞表型分化,从而促进钙、磷在骨表面沉积。
OBJECTIVE: To observe the effect of Bugu Mixture on cell differentiation by culturing bone marrow mesenchymal cells in vitro and to explore the mechanism of Bugu Mixture in treating osteoporosis. Methods: Bone marrow mesenchymal cells were isolated from SD rats. The bone marrow (BM) induced by recombinant human bone morphogenetic protein 2 (rhBMP2) was detected by morphological, alkaline phosphatase, osteocalcin, transforming growth factor β1 gene expression, Mesenchymal cells to the degree of osteoblasts to explore. Results: Bugu Mixture can promote the secretion of alkaline phosphatase and osteocalcin, and enhance the activity of rhBMP2 in differentiated bone marrow mesenchymal cells, and promote the differentiation of bone marrow mesenchymal cells to express transforming growth factor β1, This mixture of agents to promote bone marrow mesenchymal differentiation and secretory calcium is basically the same. Conclusion: Bugu Mixture can not only enhance the activity of rhBMP2, promote the differentiation of bone marrow mesenchymal cells express TGF-β, release a large amount of collagen type I in order to facilitate the deposition of calcium salts, but also promote rhBMP2-induced bone marrow mesenchymal cells Secretion of alkaline phosphatase and osteocalcin, promote bone marrow mesenchymal cells to osteoblast phenotype differentiation, thereby promoting the deposition of calcium and phosphorus in the bone surface.