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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30,60,125,250μg/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。
Objective To prepare marine sulfated polysaccharides with heparinoid structure and evaluate their in vitro anti-HBV activity using HepG2.2.15 cell model. Methods Polyglucuronic acid (PM), polyguluronic acid (PG) and chitooligosaccharides (COS), chitin (CTN) and carboxymethyl chitosan (CMC) - formamide method to prepare the corresponding polysaccharide sulfate PMS, PGS, SCOS, SCTN and SCMC, and its sulfate content, molecular weight and other physical and chemical properties were determined. The HepG2.2.15 cells were used as a model to detect the cytotoxicity of polysaccharides by MTT method. The HBsAg and HBeAg in the culture supernatant were detected by ELISA. Results HepG2.2.15 cells could inhibit the secretion of HBsAg and HBeAg at several concentrations for 30, 60, 125 and 250μg / mL for 9 days. Among them, the anti-HBV activity of PGS and PMS was better than that of three kinds of chitosan Sugar Sulfate Derivatives. Conclusion Marine heparin polysaccharides with different structures have different degrees of inhibition on HBV antigens. PGS and PMS have potential applications in anti-HBV.