九味通窍汤的体外抗脑胶质瘤作用及化学成分分析

目的研究九味通窍汤对人神经胶质瘤细胞的增殖抑制作用,并分析其化学成分。方法采用MTT法检测九味通窍汤醇提取物对人神经胶质瘤U251细胞体外增殖活性的抑制,HPLC-MS法检测其指纹图谱,并利用现有的对照品对主要成分进行结构确证。结果九味通窍汤醇提取物对U251细胞的增殖具有明显抑制作用,MTT法的剂量反应曲线为Y=0.123X+0.034 5,24h的半数抑制浓度IC50为3.78g生药/L,安全浓度IC10为0.53g生药/L。九味通窍汤的指纹图谱有28个色谱峰,含有槲皮素、毛蕊异黄酮、毛蕊异黄酮葡萄糖苷(毛蕊异黄酮-7-O-β-D葡萄糖苷)、芒柄花素、芒柄花苷(芒柄花素-7-O-β-D葡萄糖苷)。结论九味通窍汤醇提取物在体外实验中具有抗脑胶质瘤作用,应用HPLC-MS法可快速、简便、准确地测定九味通窍汤的指纹图谱,与君药的指纹图谱具有良好的相关性,并利用对照品确定了主要成分的结构,为今后研究九味通窍汤的药效成分奠定了基础。

Determination of the Inhibition Activity of Jiuwei-Tongqiao Soup on Brain Glioma in vitro and Analysis of Its Chemical Constituents

Objective To determine the proliferation inhibition activity of Jiuwei-Tongqiao soup on human glioma cells and analyze its chemical constituents.Methods The ethanol extract from Jiuwei-Tongqiao soup and its single medicinal materials were collected by reflux extraction using 95% ethanol.MTT colorimetry assay was performed to evaluate the proliferation inhibition activity of the ethanol extract from Jiuwei-Tongqiao soup on human glioma cells U251 in vitro,the fi ngerprints were detected by HPLC-MS and the structures of major components were confirmed in comparison with the reference substances.Results The proliferation of U251 cells was inhibited markedly in vitro by the ethanol extract from Jiuwei-Tongqiao soup.The dose response curve of MTT colorimetry assay is Y=0.123X＋0.034 5,the median inhibitory concentration IC50 after 24h is 3.78g/L and the safe concentration IC10 after 24h is 0.53g/L.The fingerprint of Jiuwei-Tongqiao soup has 28 peaks and contains quercetin,calycosin,calycosin-7-glucoside（calycosin-7-O-β-D-glucoside）,formononetin,and ononin（formononetin-7-O-β-D-glucoside） which belong to the monarch herba Astragalus mongholicus and Oldenlandia diffusa.Conclusion The ethanol extract from Jiuwei-Tongqiao soup has antiglioma activity in vitro.The fingerprint can be determined rapidly,conveniently and accurately by HPLC-MS and demonstrated good correlation with the fi ngerprints of the monarch harba and its structures of major components can be confi rmed in comparison with the reference substances.It lays the foundation of researching the structure of Jiuwei-Tongqiao soup＇s effective components in future.