丹参提取物促进骨髓间充质干细胞增殖及成骨分化的研究  

Salvia miltiorrhiza extract promote bone marrow mesenchymal stem cell proliferation and osteogenic differentiation

作　　者：李杨[1] 张延辉[1] 王云枫[2] 魏鑫[3] 关利新[3] 关悦[2] LI Yang ZHANG Yanhui WANG Yunfeng WEI Xin GUAN Lixin GUAN Yue(Department of Orthopedics, Hongqi Hospital of Mudanjiang Medical College, Mudanjiang, Heilongjiang 157011 Department of Endocrinology, Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011 Mudanjiang Medical University, Mudanjiang, Heilongjiang 157011)

机构地区：[1]牡丹江医学院红旗医院骨科,黑龙江牡丹江157011 [2]牡丹江医学院内分泌科,黑龙江牡丹江157011 [3]牡丹江医学院,黑龙江牡丹江157011

出　　处：《中国骨质疏松杂志》2017年第8期1030-1033,共4页Chinese Journal of Osteoporosis

摘　　要：目的研究丹参提取物对骨髓间充质干细胞增殖及成骨分化的作用。方法采用全骨髓细胞接种法和贴壁纯化培养骨髓间充质干细胞,将浓度为0.2、0.4、0.8、1.6 mg/m L和3.2 mg/m L的丹参提取物加入到骨髓间充质干细胞中,观察不同浓度的丹参提取物对骨髓间充质干细胞的增殖作用;采用0.4、0.8 mg/m L和3.2 mg/m L的丹参提取物诱导骨髓间充质干细胞向成骨细胞分化,连续诱导21 d,利用ALP试剂盒测定成骨细胞内ALP活性和茜素红染色观察成骨细胞钙化结节情况;采用Western bloting法测定成骨细胞相关蛋白BMP-2和Runx的含量。结果不同浓度的丹参提取物均能够促进骨髓间充质干细胞的增殖,丹参提取物浓度0.8 mg/m L时对骨髓间充质干细胞的增殖作用最强;ALP测定结果发现0.4、0.8 mg/m L和3.2mg/m L的丹参提取物均能够提高ALP活性,丹参提取物浓度0.8 mg/m L时ALP活性最高;茜素红染色结果发现0.4、0.8 mg/m L和3.2 mg/m L丹参提取物均能够促进成骨细胞的钙化,丹参提取物浓度为0.8 mg/m L时钙化结节数量最多;Western bloting结果表明,0.8 mg/m L的丹参提取物能够促进成骨细胞分化相关蛋白BMP-2和Runx-2表达。结论丹参提取物能够促进大鼠骨髓间充质干细胞的增殖和成骨细胞分化作用。Objective To investigate the effect of Salvia miltiorrhiza extract on the proliferation of bone marrowmesenchymal stem cells（ BMMSCs） and osteogenic differentiation. Methods BMMSCs were cultured using whole bone marrowcells inoculation method and adherent purification. Salvia miltiorrhiza extract（ SME）,in the concentrations of 0. 2,0. 4,0. 8,1. 6 mg/m L and 3. 2 mg/m L,was added to bone marrowmesenchymal stem cells and its proliferation effect on bone marrowmesenchymal stem cell were observed. Differentiation of bone marrowmesenchymal stem cells into osteoblasts had been induced by Salvia miltiorrhiza extract（ 0. 4,0. 8 mg/m L and 3. 2 mg/m L） for 21 days. ALP kit was used to determine the activity of ALP in osteoblasts and Alizarin red staining was used to observe calcified nodules in osteoblasts. The content of osteoblast related protein BMP-2 and Runx-2 was determined using Western blotting. Results Different concentrations of Salvia miltiorrhiza extract could promote the proliferation of bone marrowmesenchymal stem cells,and the strongest effect was observed for the concentration of 0. 8mg/m L. ALP assay showed that Salvia miltiorrhiza extract in the concentrations of 0. 4,0. 8 mg/m L and 3. 2 mg/m L could improve ALP activity and the concentration of 0. 8 mg/m L had the highest ALP activity. Alizarin red staining showed that Salvia miltiorrhiza extract in the concentrations of 0. 4,0. 8 mg/m L and 3. 2 mg/m L could promote bone cell calcification,and the concentration of 0. 8 mg/m L had the highest number of calcified nodules. Western blotting results showed that 0. 8 mg/m L Salvia miltiorrhiza extract could promote the expression of osteoblast differentiation related protein BMP-2 and Runx. Conclusion Salvia miltiorrhiza extract can promote the proliferation of bone marrowmesenchymal stem cells and osteogenic differentiation in rats.

关 键 词：丹参提取物 骨髓间充质干细胞 成骨分化 增殖 

分 类 号：R68[医药卫生—骨科学]