毛轴蕨多糖的分离纯化、单糖组成及对巨噬细胞RAW264.7免疫活性的影响

Purification and Monosaccharide Composition Analysis of Polysaccharides from Pteridium revolutum and Effects on Macrophage RAW264.7

目的纯化蕨菜的多糖成分,并研究其相对分子质量,单糖组成及免疫活性。方法采用热水浸提法提取毛轴蕨粗多糖,Seveg法脱蛋白,DEAE Sepharose Fast Flow和Sephadex G-200凝胶柱层析分离纯化。采用MTT法和Griess法检测了粗多糖和三种精制均一多糖对RAW264.7细胞生长和释放NO能力的影响。结果得到三种精制均一多糖组分PRP0、PRP1和PRP2;相对分子质量分别为1.04×10~6、8.39×10~5和7.37×10~5;三种精制均一多糖均含有阿拉伯糖、岩藻糖、半乳糖、甘露糖、葡萄糖、鼠李糖和木糖等单糖成分。粗多糖和三种精制均一多糖在12.5~100μg·m L^(-1)浓度范围内可显著促进RAW264.7细胞的增殖,同时均具有显著增强RAW264.7细胞释放NO的能力,并呈一定的剂量效应关系。结论蕨菜多糖可以提高免疫活性。

OBJECTIVE To purify polysaccharides from Pteridium revolutum （ Blume. ） Nakai and determine their relative molec- ular weights, monosaccharide compositions and immunomodulating activities. METHODS Crude polysaccharide was extracted from P. revolutum with hot water and then purified by DEAE Sepharose Fast Flow and Sephadex G -200 chromatography column after remo- ving proteins by Seveg method. The immunomodulating activities of the polysaccharides were investigated on macrophage cells RAW264. 7. RESULTS Three homogenous polysaccharides, PRP0, PRP1 and PRP2, were obtained from the crude polysaccharide. Their molecular weights were 1. 04 × 106, 8.39 ×105 and 7.37 × 105 , respectively. Monosaccharide composition analysis by gas chro- matography revealed that all three homogenous polysaccharides were composed of arabinose, fucose, galactose, mannose, glucose, rhamnose, and xylose in different ratios. The crude polysaccharide and three homogenous polysaccharides could promote the prolifera- tion of RAW264. 7 cells at the concentration of 12. 5 - 100 μg· mL-1 and significantly induce the production of NO from RAW264. 7 cells in a dose-dependent manner. CONCLUSION P. revolutum polysaccharides can improve immunity activity.