蛹虫草胞外多糖的提取纯化及免疫活性研究  

Study on Extraction, Purification and Immunomodulatory Effect of Cordyceps Polysaccharide of Cordyceps militaris

作　　者：王永敏[1,2] 祝文兴[2] 安利国[2] 杨桂文[2] 

机构地区：[1]山东轻工业学院食品与生物工程学院,济南250353 [2]山东师范大学生命科学学院,济南250014

出　　处：《食品与药品》2009年第2期8-11,共4页Food and Drug

基　　金：山东省自然科学基金重大项目;国家自然科学基金资助项目(No.Z2003D01)

摘　　要：目的提取纯化蛹虫草发酵液中的胞外多糖,初步研究其结构及免疫活性。方法乙醇沉淀多糖,用Sevag法除去蛋白质、透析法除去小分子物质,经CM-52纤维素柱层析后得纯品。用紫外光谱测定多糖纯度,用凝胶柱层析法测定相对分子质量,高效毛细管电泳和红外光谱分析其分子结构,检测胞外多糖对小鼠巨噬细胞系RAW264.7产生NO的影响。结果蛹虫草胞外多糖经脱蛋白、CM-52纤维素柱层析后可获得单一对称峰的纯多糖,相对分子质量为3.49×104,含羟基、糖醛结构和羰基结构,单糖基之一为甘露糖。多糖可显著刺激巨噬细胞分泌NO,具有活化免疫细胞、增强机体免疫力的作用。结论该方法可获得高纯度蛹虫草胞外多糖,并证明它对巨噬细胞有活化作用。Objective To extract and purify the cordyceps polysaccharide of Cordyceps militaris and preliminarily investigate its molecular structure and immunomodulatory effect on macrophage. Methods The pure cordyceps polysaccharide was obtained by alcohol precipitation, deproteinizing by Sevag method, removal of small molecules by dialysis and CM-52 cellulose column chromatography. The purity of cordyceps polysaccharide was determined by UV spectra; the relative molecular mass was analyzed by gel chromatography; the molecular structure was determined by high performance capillary electrophoresis and infrared spectroscopy analysis. The effect of cordyceps polysaccharide was also investigated on NO production by RAW 264.7 macrophage. Results The pure cordyceps polysaccharide with a single symmetrical peak was obtained by a series of steps of purification. Its relative molecular mass was 3.49 × 10^4 with hydroxy, furaldehyde, carbonyl groups and mannose structure. The pure cordyceps polysaccharide could stimulate RAW 264.7 macrophage to excrete NO, so it had an immunomodulatory effect on macrophage function. Conclusion The cordyceps polysaccharide of Cordyceps militaris with high purity can be produced by the above methods, and the polysaccharide can activate macrophage and improve immunity.

关 键 词：蛹虫草 胞外多糖 巨噬细胞 一氧化氮 

分 类 号：R282.71[医药卫生—中药学]