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目的探讨不同剂量健腰密骨片对小鼠椎体和骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSCs)成骨分化过程中骨形态发生蛋白-7(bone morphogenic protein-7,BMP-7)表达的影响。方法选取1月龄昆明小鼠48只,雌雄各半,按体质量随机分为健腰密骨片高、中、低剂量组及生理盐水4组,灌胃8周后取材。用Micro-CT扫描分析L4腰椎,免疫组化染色观察不同剂量健腰密骨片对椎体BMP-7表达的影响;分离培养获得各组小鼠原代骨髓间充质干细胞(BMSCs),利用碱性磷酸酶(ALP)染色、实时荧光定量RT-PCR法检测BMSCs成骨分化过程中BMP-7的表达情况。结果与生理盐水组比较,健腰密骨片高剂量组能够改善小鼠椎体三维结构,促进椎体BMP-7蛋白的表达;健腰密骨片高剂量组能够明显增加BMSCs中ALP活性,上调成骨基因BMP-7 mRNA的表达量(P<0.05)。结论健腰密骨片高剂量组能够明显提高小鼠椎体骨量,促进BMSCs成骨分化,上调椎体和BMSCs成骨分化过程中BMP-7表达,从而促进骨形成。
Objective To investigate the effect of different doses of Jianlian MiaoGu tablet on bone morphogenic protein-7 (BMP-7) during the osteogenic differentiation of mouse bone marrow mesenchymal stem cells (BMSCs) ) Expression. Methods 48 Kunming mice of 1 month old were selected and divided into three groups according to their body weight. The mice were randomly divided into four groups: high, medium and low dose of Jianhou Miangu tablet and four groups of saline. Micro-CT scanning analysis of L4 lumbar spine, immunohistochemical staining with different doses of Jianliangmige piece on the expression of BMP-7 vertebral body; isolated and cultured to obtain primary mouse bone marrow mesenchymal stem cells (BMSCs), using Alkaline phosphatase (ALP) staining and real-time fluorescent quantitative RT-PCR were used to detect the expression of BMP-7 in osteogenic differentiation of BMSCs. Results Compared with the saline group, the high dose of Jianlian Miaojian Tablet could improve the three-dimensional structure of the vertebral body and promote the expression of BMP-7 in the vertebral body. The high dose of Jianliangmiaogan Tablet could significantly increase the ALP activity, Up-regulated the expression of BMP-7 mRNA (P <0.05). Conclusion The high-dose Jian-Liang Miaowei Tablet can significantly increase the bone mass of mice, promote the osteogenic differentiation of BMSCs and up-regulate the expression of BMP-7 in the process of osteogenic differentiation of BMSCs and vertebral bodies, thereby promoting bone formation.