几种海洋类肝素多糖的制备及抗乙肝病毒活性初步研究D

目的　制备具有类肝素结构的海洋硫酸多糖,采用HepG2.2.15细胞模型评价它们的体外抗HBV活性。方法 以聚甘露糖醛酸(PM)、聚古罗糖醛酸(PG)和壳寡糖(COS)、甲壳素(CTN)及羧甲基壳聚糖(CMC)为原料,采用氯磺酸-甲酰胺法制备相应的多糖硫酸酯PMS、PGS、SCOS、SCTN和SCMC,并对其硫酸根含量、分子量等理化性质进行测定。以HepG2.2.15细胞为模型,采用MTT法检测多糖的细胞毒性,采用ELISA法检测培养上清中的HBsAg和HBeAg。结果 几种海洋类肝素多糖在30,60,125,250 μg/mL浓度下,对HepG2.2.15细胞作用9d后,均能明显抑制HBsAg和HBeAg的分泌,其中 PGS和PMS的抗HBV活性优于3种壳聚糖硫酸酯衍生物。结论　不同结构的海洋类肝素多糖对HBV抗原具有不同程度的抑制作用,PGS和PMS在抗HBV方面具有潜在的应用前景。

Preparation and antiviral activities of several marine heparin-like polysaccharides against hepatitis B virus

Objective To prepare several marine heparin polysaccharides and evaluate their activities on anti-hepatitis B virus (HBV) in vitro. Mthods Polymannuronate sulfate (PMS), polyguluronate sulfate (PGS), sulfated chitooligosaccharides (SCOS), sulfated chitin (SCTN) and sulfated carboxymethyl chitosan (SCMC) were prepared by chemical sulfation of PM, PG, COS, CTN and CMC with chlorosulfonic acid-formamide method. The contents of sulfate were determined by ion chromatography and the molecular weights were determined by HPLC. Methyl thiazolyl tetrazolium(MTT)assay was used to evaluate the cytotoxicity of the heparinoids. HBsAg and HBeAg were assessed by enzymelinked immunosorbent assay(ELISA). Results Five marine heparin-like polysaccharides exhibited considerable activities on anti-HBV in vitro. After exposed to four different concentrations of marine heparin-like polysaccharides for 9 days, the inhibition rates of extracellular HBsAg and HBeAg of each concentration increased significantly. The anti-HBV activities of PGS and PMS are better than that of chitosan sulfate derivatives. Conclusion The anti-HBV effects of marine heparin-like polysaccharides with different structures are different. PGS and PMS showed promising prospects in anti-HBV infection.