复叶耳蕨总黄酮对大鼠骨髓间充质干细胞的增殖和成骨分化的影响  

Effect of total flavonoids from Arachniodes exilis on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells

作　　者：梁广胜[1,2,3] 殷嫦嫦[1] 殷明[2] 邬亚华[1] 李辉敏[1] 刘玉亮[2,3] 魏强强[2,3] 

机构地区：[1]九江学院医学部分析检测实验室,江西九江332000 [2]南昌大学第二附属医院骨一科,江西南昌330006 [3]南昌大学研究生院医学部,江西南昌330006

出　　处：《中成药》2014年第3期456-461,共6页Chinese Traditional Patent Medicine

基　　金：采用常规实验室可用的药理研究模型系统研究复叶耳厥抑制HIV-1复制的有效成分(81160536)

摘　　要：目的研究复叶耳蕨总黄酮对大鼠骨髓间充质干细胞向成骨分化的影响。方法采用全骨髓差速贴壁法体外分离、纯化和培养大鼠骨髓间充质干细胞;取P3代细胞,分实验组和对照组,用流式细胞仪测定细胞表面标志物鉴定细胞,采用MTT法检测细胞生长曲线。显微镜下观察两组的细胞形态和组织化学染色的钙化结节;速率法测定碱性磷酸酶(ALP)活性和RT-PCR检测骨髓间充质干细胞诱导分化过程中骨桥蛋白和I型胶原的mRNA表达来鉴定成骨作用。结果体外培养的细胞表达表面标志;复叶耳蕨总黄酮对骨髓间充质干细胞内ALP活性增高并形成明显钙结节;PCR结果显示复叶耳蕨总黄酮上调骨桥蛋白和I型胶原的mRNA表达。结论 20μg/mL复叶耳蕨总黄酮可促进大鼠骨髓间充质干细胞定向分化为成骨样细胞。AIM To study the effect of total flavonoids from Arachniodes exilis on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells （BMSCs）. METHODS Rat BMSCs, isolated from rats bone marrow in vitro by whole bone marrow adherent method, was purified and cultured. Identification of thesurface marker of BMSCs and determination of their growth curve were completed by MTF. They were divided into experimental （total flavonoids from Arachniodes exilis） and control groups, and osteogenesis was identified by mi- croscopic cell morphology, the activity of alkaline phosphatase , the formation of calcified nodules by staining, and the expression of osteopontin and collagen 1 were tested by RT-PCR. RESULTS Cells cultured in vitro expressed the surface markers of BMSCs. The total flavonoids from Arachniodes exilis improved the activity of ALP, promoted the formation of calcium nodule, and increased the expression of osteopontin and I collagen shown by PCR results. CONCLUSION 20μg/mL total flavonoids from Arachnides exilis can promote rat bone marrow mesenchymal stem cells to differentiate into bone cells.

关 键 词：复叶耳蕨总黄酮 骨髓间充质干细胞 增殖 碱性磷酸酶 成骨分化 

分 类 号：R285.5[医药卫生—中药学]