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  

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

作　　者：唐德志[1] 杨洲[1] 笪巍伟[1] 赵永见[1] 姚长风[1] 施杞[1] 王拥军[1] 

机构地区：[1]上海中医药大学附属龙华医院,上海中医药大学脊柱病研究所,上海200032

出　　处：《上海中医药杂志》2016年第7期72-75,93,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：科技部国家重点基础研究发展计划(973计划)资助项目(2010CB530400);国家自然科学基金面上项目(81473701);上海市科委青年科技启明星计划项目(14QA1403500);上海市科委中药领域科技支撑项目(13401900304);上海市科委中医重点项目(15401971700);上海市卫计委新优青培养计划项目(XYQ2013085);上海市教委科研创新项目(14YZ051)

摘　　要：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 study the effect of different doses of JianYao-MiGu tablet on mice bone formation and BMP7 expression nf bone marrow mesemnehymal stem cells （BMSCs） in mice during the osteogenetie differentiation prncess. Methods Sixty l-month-old Kunming mice, male thirty and female thirty, were randomly divided into four grnups of high-dose, middle-dose, low-dose, aud saline group. A II 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 immunohistoehemieal staining. Primary hone 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 PCPt analysis was performed to observe the mRNA expression of BMP-7. Results Compared to norrnal saline group, high-dose group improved mice vertebral three-dimensiooal structure, increased the activity of ALP, prc）moted the mRNA anti protein expression of BMP-7 expression. Conclusion High-dose JianYao-MiGu tablet could promote the express/on of ALP in BMSCs and osteogenic differentiation, increase the expressinn of BMP-7, thus as to promote bone fo formation.

关 键 词：健腰密骨片 骨髓间充质干细胞 骨形态发生蛋白-7 骨形成 中医药疗法 

分 类 号：R285.5[医药卫生—中药学]