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利用DEAE-Sepharose fast flow色谱层析柱及真空冻干技术对栗黑褐拟层孔菌胞外多糖进行分离纯化,并利用苯酚-硫酸法、红外光谱、高效液相色谱等化学分析手段对主要多糖进行结构表征;通过构建小鼠模型,对小鼠脏器系数进行分析,提取回肠及空肠组织RNA,采用Real-time RT-PCR方法对免疫相关基因IL-4、β-actin、IFN-γ和TNF-α的表达情况进行分析。研究表明:栗黑褐拟层孔菌菌丝体内多糖主要由鼠李糖、阿拉伯糖、甘露糖、葡萄糖及半乳糖组成,其摩尔比为43.40:0.09:0.13:51.32:5.06;高效液相色谱计算主要多糖分子量为3.24×104 u,红外光谱分析表明栗黑褐拟层孔菌胞外多糖具有多糖类物质的结构特征;相关免疫脏器系数随着天数的增加而增加,实验组的脏器系数较对照组相比,显著增加;相关免疫调控基因IL-4、β-actin、IFN-γ和TNF-α的表达量随着时间的延长均有所增加。
The extracellular polysaccharides of Porphyra yezoensis were isolated and purified by DEAE-Sepharose fast flow chromatography and vacuum freeze-drying technique. The chemical constituents such as phenol-sulfuric acid, infrared spectroscopy and high performance liquid chromatography Polysaccharides were characterized by real-time RT-PCR. The expression of IL-4, β-actin and IFN-γ in the intestine of ileum and jejunum tissues were analyzed by constructing a mouse model. And TNF-α expression analysis. The results showed that the polysaccharides in Mycelium of M. avium were mainly composed of rhamnose, arabinose, mannose, glucose and galactose with the molar ratio of 43.40: 0.09: 0.13: 51.32: 5.06, and high performance liquid chromatography The molecular weight of the main polysaccharide was calculated to be 3.24 × 104 u. Infrared spectrum analysis showed that polysaccharide had the structural characteristics of polysaccharide. The relative immune organ coefficient increased with the increase of days, The coefficient of IL-4, β-actin, IFN-γ and TNF-α were increased with time compared with the control group.