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目的寻找和筛选具有较好佐剂功能的天然多糖。方法自中药提取当归多糖、黄芪多糖、板蓝根多糖、茯苓多糖、黄精多糖、枸杞多糖和淫羊藿多糖。苯酚硫酸法测总糖含量;间羟联苯法分析糖醛酸含量;凝胶渗透色谱法测定多糖分子量分布。按照分组,每只BALB/c小鼠肌内注射60μg卵清蛋白(OVA),OVA+0.1mg氢氧化铝,OVA+1mg多糖,OVA+生理盐水,并分别在第1,28和58天进行3次注射,在第21,52,70天采血,测定血清OVA抗体滴度。结果 7种多糖中的总糖含量为板蓝根总糖,65.41%;当归总糖,30.88%;黄芪总糖,43.70%;黄精总糖,48.88%;茯苓总糖,58.68%;枸杞总糖,45.83%;淫羊藿总糖,32.60%;总糖中糖醛酸含量为板蓝根糖醛酸,13.36%;当归糖醛酸,19.73%;黄芪糖醛酸,6.53%;黄精糖醛酸,5.96%;茯苓糖醛酸,1.96%;枸杞糖醛酸,8.96%;淫羊藿糖醛酸,7.53%。7种多糖表现出不同的分子质量分布特征。初次免疫,7种多糖和铝佐剂均未激活小鼠血清OVA抗体产生;在第2次免疫后,淫羊藿多糖佐剂组产生较高滴度的抗体,达到1:105;第3次免疫后淫羊藿多糖佐剂组抗体滴度进一步提高,板蓝根多糖、当归多糖及茯苓多糖佐剂组的OVA特异性抗体滴度均达到1:105,与OVA+0.1mg氢氧化铝及OVA+生理盐水组比较有显著差异(P<0.05)。结论板蓝根多糖、当归多糖、茯苓多糖及淫羊藿多糖均具有很好的佐剂作用,特别是淫羊藿多糖具有较强的激发体液免疫活性。多糖有望成为候选的新型免疫佐剂。
Objective To search and screen natural polysaccharide with good adjuvant function. Methods Angelica sinensis polysaccharide, Astragalus polysaccharide, Radix isatidis, tuckahoe polysaccharide, Polygonatum polysaccharide, Lycium barbarum polysaccharide and Epimedium polysaccharides were extracted from the Chinese medicine. Determination of total sugar content by phenol sulfuric acid method; Analysis of uronic acid content by m-hydroxybiphenyl method; Molecular weight distribution of polysaccharide by gel permeation chromatography. Each BALB / c mouse was intramuscularly injected with 60 μg of ovalbumin (OVA), OVA + 0.1 mg of aluminum hydroxide, OVA + 1 mg of polysaccharide, OVA + saline, and was administered on days 1, 28 and 58, respectively On the 21st, 52th and 70th day, blood samples were collected for determination of serum OVA antibody titers. The results showed that the total sugar content in the seven polysaccharides was 65.41%, total sugar in Angelica sinensis, 30.88%, total sugar in Astragalus, 43.70%, total flavonoids in Huangjing, 48.88%, total sugar in Poria cocos, 58.68% %; Epimedium total sugar, 32.60%; the total sugar uronic acid content is Radix isoniazid, 13.36%; Angelica ursolic acid, 19.73%; Astragaluronic acid, 6.53% ; Poriacuronic acid, 1.96%; wolfberry uronic acid, 8.96%; Epimedium uronic acid, 7.53%. Seven kinds of polysaccharides showed different molecular mass distribution characteristics. Primary immunization, seven kinds of polysaccharides and aluminum adjuvant did not activate serum OVA antibody in mice; after the second immunization, Epimedium polysaccharide adjuvant group produced higher titer antibody, reaching 1: 105; the third After immunization Epimedium polysaccharide adjuvant group antibody titers further increased, the Radix isatidis polysaccharide, Angelica polysaccharide and Poria polysaccharide adjuvant group OVA specific antibody titer reached 1: 105, and OVA +0.1 mg of aluminum hydroxide and OVA + physiology There was significant difference between saline group (P <0.05). Conclusion Radix isatidis polysaccharide, Angelica sinensis polysaccharide, tuckahoe polysaccharides and Epimedium polysaccharides have a very good adjuvant effect, especially Epimedium polysaccharides have a strong stimulated humoral immune activity. Polysaccharide is expected to become a candidate of a new immune adjuvant.