牛膝中脱皮甾酮的含量测定及促成骨样细胞增殖活性

牛膝为苋科牛膝属植物ＡｃｈｙｒａｎｔｈｅｓｂｉｄｅｎｔａｔａＢｌ.之干燥根 ,有散淤血、消肿痛、补肝肾、强筋骨之功效[1] ,为常用中药之一。历代文献明确为治疗“骨痹”、“骨痿”(骨质疏松症 )的方剂共 2 1个 ,涉及药物 77种 ,其中牛膝是出现频率较高的药物[2 ] 。通过研究发现牛膝的醇提物以及醇提物的乙酸乙酯萃取物对成骨样细胞ＵＭＲ10 6有显著的促进增殖作用[3] 。因此 ,我们以成骨样细胞ＵＭＲ10 6的增殖为靶向指标 ,对牛膝醇提物的乙酸乙酯萃取物再进行提取、分离、纯化 ,找出较强的促进细胞增殖的活性成分 ,并研究ＨＰＬＣ含…

DETERMINATION OF ECDYSTERONE IN ACHYRANTHES BIDENTATA BL. AND ITS ACTIVITY PROMOTING PROLIFERATION OF OSTEOBLAST-LIKE CELLS

AIM: To study the activity of ecdysterone from Achyranthes bidentata Bl. (AB) promoting proliferation of osteoblast-like (OB-like) UMR106 cells and to determine its content in AB by HPLC method. METHODS: Ecdysterone isolated from AB was cultured with OB-like cells UMR106 together in vitro and the proliferation of OB-like cells was determined by MTT assay. The chromatographic conditions for determining ecdysterone included an ODS column (250 mm x 4.6 mm, 5 microns), a mobile phase consisting of a mixture of water-acetontrile-tetrahydrofuran (86:11:3), detection wavelength of 243 nm, and column temperature of 27 degrees C. Phenacetin was used as the internal standard. RESULTS: The ecdysterone from AB had significant activity promoting proliferation of OB-like cells, the proliferation was promoted by 41% (n = 3). The average recovery of ecdysterone was 96.2% (RSD = 2.1%), the calibration was linear in the range of 30-300 micrograms.mL-1 (gamma = 0.9998). CONCLUSION: Ecdysterone was screened quickly by cultivating with OB-like cells together in vitro. The HPLC method is accurate, fast and reproducible for the determination of ecdysterone in AB.