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目的:研究健腰密骨片对骨形成及骨髓间充质干细胞(BMSCs)骨形态发生蛋白(BMP-7)、骨钙素(OC)表达的影响。方法:选取1月龄昆明小鼠60只,雌雄各半,按体质量随机分为健腰密骨片高、中、低剂量组,仙灵骨葆组,生理盐水组,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、OC表达的影响;分离培养获得各组小鼠原代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、OC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,明显升高小鼠椎体骨密度(P<0.05),增加椎体骨小梁数目和骨小梁厚度增加(P<0.05),降低骨小梁分离度(P<0.05),促进椎体BMP-7、OC蛋白的表达;健腰密骨片高剂量组能够明显促进BMSCs中ALP的表达,上调成骨基因BMP-7、OC m RNA的表达量,其中OC m RNA升高显著(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密度,促进BMSCs成骨分化,上调椎体和BMSCs成骨分化过程中BMP-7、OC表达,从而促进骨形成。
Objective: To study the effect of Jianlian MiaoGu Tablet on bone formation and bone morphogenetic protein (BMP-7) and osteocalcin (OC) expression of bone marrow mesenchymal stem cells (BMSCs). Methods: Sixty Kunming mice of 1 month old were randomly divided into three groups: male, female, male, female, male, female, male and female. One week old Kunming mice were randomly divided into high, medium and low dose group, Xianlinggubao group and normal saline group Drawn. Micro-CT scanning analysis of L4 lumbar spine, immunohistochemical staining with different doses of Jianliangmige piece on the expression of BMP-7, OC; isolated and cultured to obtain the primary mouse BMSCs in each group, the use of alkaline phosphatase ( ALP) staining. Real-time fluorescent quantitative RT-PCR was used to detect the expression of BMP-7 and OC in osteogenic differentiation of BMSCs. Results: Compared with the saline group, the high-dose Jian-Liang Miaowei tablet group could improve the three-dimensional structure of the vertebral body and significantly increase the vertebral bone density (P <0.05), and increase the number of trabecular bone and the small bone (P <0.05), reduce the trabecular separation (P <0.05), and promote the expression of BMP-7 and OC protein in vertebral body. The high dose of Jianliangmigeng tablet can obviously promote the expression of ALP in BMSCs, Up-regulated the expression of osteoblast BMP-7 and OC m RNA, and the OC m RNA increased significantly (P <0.05). Conclusion: The high-dose Jianlianmiou tablet group can significantly increase the BMD of mice, promote the osteogenic differentiation of BMSCs and up-regulate the expression of BMP-7 and OC during the osteogenic differentiation of vertebral body and BMSCs, and promote bone formation.