白术多糖的免疫佐剂作用

采取水提醇沉、Sevage法除蛋白、D101大孔树脂柱分离、透析等方法提取纯化白术多糖,用苯酚硫酸法测定其多糖含量;白术多糖经三氟乙酸高温水解,醋酸酐乙酰化后进行气相色谱分析其单糖组成。将35只ICR小鼠随机分成7组,每组5只。以卵清白蛋白（OVA）为模式抗原,分别设含150、300或600μg白术多糖或黄芪多糖组,对照组仅免疫OVA。抗原和佐剂混合均匀后,腹部皮下注射疫苗,共免疫2次,间隔2周。二次免疫后2周摘除眼球采血,间接ELISA法检测血清抗OVA抗体及亚类水平。结果显示,白术多糖得率为4.2%,多糖含量为86.2%,由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖组成;OVA疫苗中添加白术多糖能显著提高ICR小鼠血清中抗OVA特异性抗体及抗体亚类水平,表明白术多糖皮下注射后具有免疫佐剂作用。

Immunological adjuvant effect of polysaccharide from Rhizoma Atractylodis Macrocephalae

In experiment A,Rhizoma Atractylodis Macrocephalae polysaccharides（RAMPS）were extracted with boiling water and precipitated by 95% ethanol,after removal of protein by Sevage method,the polysaccharide were further purified by Macroporous Adsorption Resin column D101 and dialyzed against distilled water.Total sugar content was estimated by the phenol-sulfuric acid analysis using glucose as a standard.The polysaccharides were hydrolyzed with TFA and derivatized to acetylated aldononitriles and isothermally separated by gas chromatography（GC）to analyse monosaccharides component.In experiment B,thirty-five female ICR mice were randomly divided into five groups with 7 mice in each.Mice were immunized with OVA alone as control,or in combination with 150,300 or 600 μg RAMPS or Astragalus polysaccharides.Two weeks after the first immunization,mice were boosted with the same route.Blood samples were then collected two weeks after the boost for detection of serum OVA-specific antibody and subclasses.The results indicated that the yield of RAMPS in this experiment was 4.2% with polysaccharides content of 86.2%,which was composed of rhamnose,arabinose,xylose,mannose,glucose and galactose.RAMPS could significantly improve the level of serum specific IgG and subclass response of mice,suggesting the immunological adjuvant effect of RAMPS.