目的观察淫羊藿中单体黄酮的抗骨质疏松作用。方法利用硅胶和凝胶柱色谱分离淫羊藿中单体黄酮成分,根据化合物光谱数据(ESI-MS、1H-NMR和13C-NMR)鉴定其结构,通过MTT法观察其对体外培养的前破骨细胞株RAW264.7增殖的影响。结果从淫羊藿地上部分的醋酸乙酯萃取物中分离得到5个化合物,分别为淫羊藿苷()、宝藿苷()、朝藿素B()、宝藿苷()及金丝桃苷()。活性结果表明,0.1~100μmol/L化合物对前破骨细胞RAW264.7的生长有一定程度的促进作用;浓度为0.1~100μmol/L的化合物对前破骨细胞RAW264.7的生长有一定的抑制作用,但随着浓度的升高,转而表现为轻微的促进作用;浓度为1.0和10μmol/L的化合物对该细胞的增殖基本上没有什么影响,而其他浓度下,则对前破骨细胞RAW264.7的增殖表现出明显的抑制作用;除个别浓度外,化合物和均对前破骨细胞RAW264.7的生长表现出相当程度的抑制作用。结论黄酮类化合物对RAW264.7的生长是促进还是抑制依赖于其自身的化学结构以及作用浓度,淫羊藿可能是通过黄酮类化合物抑制前体破骨细胞的增殖,进而抑制前体破骨细胞分化形成破骨细胞而发挥抗骨质疏松作用的。

Objective To study the antiosteoporosis activity of flavonoids isolated from Epimedium koreanum. Methods The compounds were separated by column chromatography with silica gel and Sephadex LH-20 and identified by spectral analysis (ESI-MS, ~1H-NMR , and ~ 13 C-NMR ), respectively. MTT Assay was used to investigate the effects of the compounds on proliferation of preosteoclast RAW 264.7 cell line in vitro. Results Five compounds were isolated from the ethyl acetate extract of the aerial part of E. koreanum. Their structures were identified as icariin (Ⅰ), baohuoside Ⅱ (Ⅱ), epimedokoreanin B (Ⅲ), baohuoside Ⅰ (Ⅳ), and hyperoside (Ⅴ). The results indicated that compound Ⅱ could promote the proliferation of RAW 264.7 cell line at concentrations of 0.1-100 μmol/L. Compound Ⅰ at concentrations of 0.1-100 μmol/L could inhibit the proliferation of RAW 264.7 cell line and turned to stimulate the proliferation of RAW 264.7 cell line as concentration increased. Compound Ⅲ at concentrations of 1.0 and 10 μmol/L had no effect on the proliferation, but significantly inhibited the proliferation of RAW 264.7 cell Line at other concentrations. Except individual concentration, compounds Ⅳ and Ⅴ at concentrations of 0.1-100 μmol/L significantly inhibited the proliferation of RAW 264.7 cell line. Conclusion The effects of flavonoids on the proliferation of RAW 264.7 cell line are bi-directional, depending on concentrations and their chemical structures. The osteoporsis may be controlled by E. koreanum through inhibiting the proliferation and differentiation of preosteoclast.

国家863计划资助项目(2003AA2Z2052)