目的:对不同提取方法所得的卷柏提取物进行雌激素活性筛选,探索卷柏发挥雌激素作用的活性部位。方法:采用小鼠子宫增重试验,将卷柏分别以70%乙醇回流所得总部位经聚酰胺柱分离;95%乙醇回流药渣经水提取;药渣水提物去除多糖等所得的卷柏各组分进行雌激素活性筛选。运用HPLC指纹图谱技术对卷柏95%醇提物、药渣水提物进行分析。结果:70%乙醇回流所得的卷柏总部位、药渣水提物高剂量、药渣水提物去多糖组与空白组比较,均能显著性增加小鼠子宫系数(P〈0.01),卷柏其余各组与空白组相比无明显作用;HPLC指纹图谱显示卷柏95%醇提物中穗花杉双黄酮为其主要成分,而药渣水提物中基本不含穗花杉双黄酮。结论:95%乙醇回流药渣水提法与聚酰胺柱分离法相比,前者更适合于卷柏雌激素活性物质的提取分离;水溶性除多糖物质为卷柏中发挥雌激素样作用的有效部位;HPLC指纹图谱结果提示,穗花杉双黄酮不是卷柏发挥雌激素作用的成分。
Objective:To identify the suitable extraction method and polarity for estrogenic active substance in Selaginella tamariscina,through the estrogenic active screening of different extracts of S.tamariscina from different extraction methods.Method:Selaginella tamariscina was extracted by 70% ethanol to obtain the total fraction.The total fraction was separated by polyamide column to obtain the water fraction,50% ethanol and 80% ethanol fractions;S.tamariscina was refluxed by 95% ethanol to obtain the 95% ethanol extract,and the residue was decocted by water to obtain the residue water extract;the residue water extract was removed polysaccharides to obtain non-polysaccharides part of water-soluble components.Then the uterine weight method in mice was adopted to detect the estrogenic activity of all these different extracts.The 95% ethanol extract and the residue of water extract were analyzed by high-performance liquid chromatography(HPLC) fingerprint.Result:Compared with the control group,the total fraction,high dose of the water extract residue and non-polysaccharides part of water-soluble components group significantly increased uterus coefficient(P0.01),while the other groups of S.tamariscina showed no significantly effects(P0.05);HPLC fingerprint demonstrated that amentoflavone was the main component of the 95% ethanol extract,but no amentoflavone was observed in the residue water extract.Conclusion:The extraction method of 95% ethanol refluxing and the residue water decocting was more suitable for extracting the estrogenic active substance of S.tamariscina;the non-polysaccharides part of water-soluble components played an important role in estrogen activity in S.tamariscina;HPLC fingerprint suggested that amentoflavone was not the estrogenic active component of S.tamariscina.