微生物多糖183的分离纯化和抗乙肝病毒活性研究

Isolation and anti-HBV activities of microbial polysaccharides 183

目的研究链霉菌183发酵液中抗乙型肝炎病毒活性成分。方法通过超滤法和凝胶过滤层析进行分离纯化,用高效凝胶渗透法和多角度激光光散射法测定分子量,用酸水解、糖醇乙酸酯衍生化和气相色谱测定单糖组成和摩尔比,用鸭乙肝病毒复制复合体DNA聚合酶/逆转录酶筛选模型,2.2.15细胞和鸭乙型肝炎病毒动物模型评价抗乙肝病毒活性。结果分离得到183A、B、C、D四个多糖纯品,183B在2.2.15细胞内抑制HBV DNA的IC50为2.62μg/mL,粗多糖在接近鸭最大耐受剂量时对鸭乙肝病毒显示一定的抑制作用。结论在酶水平和细胞水平上的两个不同层次的模型中,183各多糖组分的抗病毒效果并不平行,183粗多糖对病毒复制关键酶及2.2.15细胞内HBV复制的抑制作用较强。

Objective To study the active anti-HBV components from Streptomyces sp. 183. Methods Ultrafiltration and gel filtration chromatography were used to separate active polysaccharides. Molecular weights were measured by HPGPC and MALLS. Composition and molar ratio of monosaccharides were determined by acid hydrolysis, alditol acetate derivative method and GC. Anti-HBV activities were evaluated by duck HBV RCs DNAP/ RT model, 2.2.15 cell and DHBV animal model. Results Four pure polysaccharides, 183A, B, C and D were isolated. 183B showed strong inhibitory effect against HBV DNA in 2.2.15 cell. Crude polysaccharides showed some inhibitory effect against DHBV replication in infected ducks. Conclusion Crude polysaccharides showed strong inhibitory effects against key enzyme in DHBV replication and HBV DNA in 2.2.15 cell. The anti-HBV activities of four pure polysaccharides were not parallel in enzyme level and cell level models.