黄边灵芝多糖的结构鉴定及免疫活性的研究

Study on the structure identification and immune activity of Ganoderma luteomarginatum polysaccharide

以四川小金县野生黄边灵芝为研究材料,采用热水浸提法、乙醇醇沉法对黄边灵芝中的多糖进行提取,并通过DEAE-cellulose column柱层析等方法对多糖进行分离纯化,采用高效凝胶渗透色谱(HPGPC),红外光谱(FTIR),核磁共振谱(1H NMR和13 C NMR)等技术对黄边灵芝多糖的结构进行初步鉴定,并用CCK-8法在体外对其免疫活性进行测定。结构鉴定结果显示,黄边灵芝多糖的分子量约为7.96×10^3 Da,由3种单糖所组成,具有α-D-吡喃糖主链,β-D-吡喃糖侧链结构,主链与侧链比为4∶1,结果表明黄边灵芝多糖是一种结构新颖的均一多糖。免疫活性结果显示,在浓度梯度为1.25~20μg/mL的黄边灵芝多糖作用下,T细胞、B细胞、RAW264.7巨噬细胞与空白对照组相比,OD值增加,细胞明显成团。说明在一定浓度范围内,黄边灵芝多糖与空白对照组相比能够极显著(P<0.01)的促进T细胞、B细胞、RAW264.7细胞的增殖。

Taking wild Ganoderma luteomarginatum in Xiaojin County of Sichuan as the research material,the polysaccharides were extracted by hot water extraction and ethanol alcohol precipitation method.The polysaccharides were separated and purified by DEAE cellulose column chromatography and so on.High performance gel permeation chromatography(HPGPC),infrared spectroscopy(FTIR)and nuclear magnetic resonance spectroscopy(1H NMR and 13 C NMR)were used.The structure of G.luteomarginatum polysaccharide was preliminarily identified and its immunological activity was determined by CCK-8 method in vitro.The results of structure identification showed that the molecular weight of G.luteomarginatum polysaccharide was about 7.96×10^3 Da,which was composed of three kinds of monosaccharides.It had the structure ofα-D-pyranose main chain andβ-D-pyranose side chain.The ratio of main chain to side chain was 4∶1.The results showed that G.luteomarginatum polysaccharide was a kind of uniform polysaccharide with novel structure.The results of immunological activity showed that under the action of G.luteomarginatum polysaccharide with concentration gradient of 1.25~20μg/mL,the OD value of T cells,B cells and RAW264.7 macrophages increased and the cells were obviously clustered compared with the blank controlled group.The results showed that G.luteomarginatum polysaccharide could significantly promote the prol iferation of T cells,B cells and RAW264.7 cells in a certain concentration range compared with the blank controlled group(P<0.01).