摘要
目的研究淫羊藿苷对体外培养骨髓基质干细胞增殖与成骨性分化的影响,了解其是否具有促进骨形成活性。方法将10-8、10-7、10-6、10-5和10-4mol/L 5种浓度的淫羊藿苷分别加入原代培养大鼠骨髓基质干细胞(rBMSCs)中,采用MTT法分析细胞增殖情况。在成骨性诱导培养条件下,比较淫羊藿苷组和不加药的对照组间在碱性磷酸酶阳性克隆数(CFU-FALP)、碱性磷酸酶(ALP)活性、骨钙素分泌量、钙盐沉积量和钙化结节数等方面的差异,采用RT-PCR和实时荧光定量PCR分析成骨性分化的标志蛋白和细胞因子的基因表达差异。结果淫羊藿苷无促进细胞增殖活性,但10-5mol/L及更高浓度抑制细胞增殖。10-5mol/L淫羊藿苷强烈促进原代培养rBMSCs的成骨性分化,表现在加药组的CFU-FALP数量、胞内ALP活性、骨钙素分泌量、钙盐沉积量和钙化结节数等在培养不同阶段均高于或显著高于对照组,包括骨涎蛋白、骨桥素、类胰岛素-I和BMP2等在内的多种标志蛋白和细胞因子基因表达水平也显著提高。结论淫羊藿苷强烈促进骨髓基质干细胞的成骨性分化,表明其具有促进骨形成生理活性。
Objective To investigate the effects of icariin on the osteogenic differentiation of bone marrow mysenchymal stem cells. Methods 10^-8 , 10^- 7, 10^- 6, 10^- 5 and 10^-4 mol/L icariin were supplemented into the culture media of primary rat bone marrow mysenchymal stem cells （rBMSCs） separately. The cell proliferation was assayed by MTT method and compared with each other. The osteogenic differentiation markers including CFU-FALP , alkaline phosphatase （ALP） activity, osteocalcin secretion, calcium deposition and mineralized bone modulus were compared between the icariin-supplemented group and the control. The gene expression of bone-related protein and cytokines were analysed by common RT-PCR and real-time RT-PCR. Results Icariin didn＇t stimulate but inhibit the proliferation of rBMSCs at primary culture. 10^-5 mol/L icariin strongly enhances the osteogenic differentiation of rBMSCs which was shown by significantly improved CFU-FALP number, ALP activity, osteocalein secretion, calcium deposition and mineralized modulus in the icarrin-supplemented group compared with the control. The gene expression level of bone-related marker proteins and cytokines were also obviously higher in the former than in the latter group. Conclusion Icariiu strongly stimulates the osteogenic differentiation of BMSCs, which indicates that icariin has the ability to enhance the bone formation activity.
出处
《中国骨质疏松杂志》
CAS
CSCD
2008年第9期642-645,632,共5页
Chinese Journal of Osteoporosis
基金
全军医学科学技术重点资助项目(01Q012)
中国博士后科学基金资助项目(2005037575)
关键词
淫羊藿苷
骨髓基质干细胞
成骨性分化
增殖
Icariin
Mysenchymal stem cells
Osteogenic differentiation
Proliferation
