不同剂量健腰密骨片对骨形成和小鼠骨髓间充质干细胞中BMP-7表达的影响

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(P0.05)。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。

The effect of different dose JianYao-MiGu tablet on mice bone formation and BMP 7 Expression of bone marrow mesenchymal stem cells

  Objective:To study the effect of different doses of JianYao-MiGu tablet on mice bone formation and BMP7 expression of bone marrow mesemnchymal stem cells (BMSCs) in mice during the osteogenetic differentiation process. Methods Sixty 1-month-old Kunming mice, male thirty and female thirty, were randomly divided into four groups of high-dose, middle-dose, low-dose, and saline group. All the mice were treated intragastrically and sacrificed after 8 weeks. The fourth lumbar vertebra (L4) was scanned by Micro-CT, The BMP-7 expression of each group were observed using immunohistochemical staining. Primary bone marrow-derived mesenchymal stem cells (BMSCs) were isolated and cultured in each group. Alkaline phosphatase (ALP) staining was performed to observe the osteogenic differentiation, and real-time quantitative PCR analysis was performed to observe the mRNA expression of BMP-7.  Results: Compared to normal saline group, high-dose group improved mice vertebral three-dimensional structure, increased the activity of ALP, promoted the mRNA and protein expression of BMP-7 expression.  Conclusion:High-dose JianYao-MiGu tablet could promote the expression of ALP in BMSCs and osteogenic differentiation, increase the expression of BMP-7, thus as to promote bone formation.