Gao X Y, Wang D W, Li F M
Shenyang Pharmaceutical University, Shenyang 110015, China.
Yao Xue Xue Bao. 2000 Nov;35(11):868-70.
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.
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.
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).
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.
研究牛膝中蜕皮甾酮促进成骨样（OB样）UMR106细胞增殖的活性，并采用高效液相色谱法测定其在牛膝中的含量。
将从牛膝中分离得到的蜕皮甾酮与OB样细胞UMR106进行体外共培养，采用MTT法测定OB样细胞的增殖情况。测定蜕皮甾酮的色谱条件包括：ODS柱（250mm×4.6mm，5μm），流动相为水-乙腈-四氢呋喃（86:11:3）混合液，检测波长为243nm，柱温为27℃。以非那西丁为内标。
牛膝中的蜕皮甾酮具有显著促进OB样细胞增殖的活性，增殖率提高了41%（n=3）。蜕皮甾酮的平均回收率为96.2%（相对标准偏差=2.1%），在30-300μg·mL-1范围内校准曲线呈线性（γ=0.9998）。
通过与OB样细胞体外共培养可快速筛选出蜕皮甾酮。高效液相色谱法测定牛膝中蜕皮甾酮准确、快速且重现性好。
