【作者】 杨涛；

【导师】 封士兰；

【作者基本信息】 兰州大学 ， 药物分析， 2014， 硕士

【摘要】 本文主要研究了红芪经水提30%乙醇沉淀得到的红芪多糖1(HPS1)及从HPS1中分离出的四个组分,HPS1-A、HPS1-B、HPS1-C、HPS1-D的理化性质、结构特征和抗补体活性。HPS1经过除蛋白、脱色素、DE-52阴离子交换柱分离纯化得到一均一组分HPS1-C和混合部分(HPS1-1),HPS1-1经凝胶SephadexG-100柱色谱进一步分离纯化得到三个均一组分,HPS1-A、HPS1-B、HPS1-D。这四部分的理化性质和构象进行了研究,除此之外还通过GPC-MALLS法和扫描电镜(SEM)研究了多糖的构象采用部分酸水解,糖醛酸还原,甲基化,气相色谱,气质联用(GC-MS),红外光谱,多角度激光散射仪及一维、二维核磁共振波谱,研究了红芪多糖HPS1-B和HPS1-D的结构。结果显示HPS1-B是一种酸性多糖,分子量523.6KDa,主链是由α-L-Araf(1→,→5)α-L-Araf(1→,β-D-Glcp(1→,→4)α-D-Glcp(1→,→4,6)α-D-Glcp(1→组成,侧链由→4)α-D-GlapA(1→,β-D-Glcp(1→和→4)α-D-Glcp(1→组成,并且含有硫酸基,每隔18-22个残基重复出现一次。构象研究表明,HPS1-B在0.9%NaCl和质量分数为0.2%叠氮化钠溶液溶液中呈无规则线团状。对HPS1-D的结构研究同HPS1-B方法一致。结果显示,HPS1-D是一种中性杂多糖,不含硫酸基,分子量为45.93KDa,主链骨架由→4)α-D-Glcp(1→,→4,6)α-D-Glcp(1→组成,侧链分支位于葡萄糖O-6位。侧链分支主要由α-L-Araf(1→,→5)α-L-Araf(1→,→3,5)α-L-Araf(1→组成,可能还有少量的1,4连接的葡萄糖存在。主链末端为葡萄糖,侧链末端主要由阿拉伯糖组成,可能有少量的葡萄糖。其构象在0.9%NaCl和质量分数为0.2%叠氮化钠溶液中以无规则线团状的形式存在。采用经典途径的方法对红芪多糖HPS1、HPS1-A、HPS1-B、HPS1-C和HPS1-D的体外抗补体活性进行了研究。结果显示在经典途径下HPS1-D、 HPS1-A、HPS1-B抗补体活性较好,HPS1次之,HPS1-C的活性较弱。更多还原

【Abstract】 Four polysaccharides, HPS1-A, HPS1-B, HPS1-C and HPS1-D, were obtained by aqueous extraction followed by precipitation with ethanol and purified with anion-exchange on DEAE-52cellulose and gel-permeation chromatography on sephadex G-100from Hedysarum polybotrys Hand.-Mazz.In this paper we studied the structural features and anti-complementary activity of above-mentioned four fractions and Radix Hedysari polysaccharides1(HPS1). We compared the physicochemical properties of above-mentioned four fractions by the total sugar content, protein content, uronic acid content, elements composition, and molecular weight, monosacch-aride composition. In addition, we also compared the conformation of HPS1-A, HPS1-B, HPS1-C and HPS1-D by high performance gel permeation chromatography-multi angle laser light scattering (GPC-MALLS) and scanning electron microscopy(SEM).Then partial acid hydrolysis,uronic acid reduction, methylation, GC, GC-MS, IR, multi-angle laser light scattering and one-dimensional and two-dimensional nuclear magnetic resonancespectroscopy method were used to study chemical structural of HPS1-B and HPS1-D. The result showed that HPS1-B was a acidic polysaccharide, molecular weight was measured to be523.6KDa, which backbone was composed of a-L-Araf (1→,→5) α-L-Araf (1→β-D-Glcp (1→,→4)α-D-Glcp(1→,→4,6) α-D-Glcp (1→, residues substituted at0-6of galacturonopyranose. And that the side chains were composed of→4)α-D-GlapA(1→,β-D-Glcp (1→and→4) α-D-Glcp(1→), and occasionally by SO3-, and one sulfate ester occasionally occur per18-22residues. Its conformation has been studied by GPC-MALLS, the results showed that the conformation of HPSl-B has a random coil, and was mono-dispersed in0.9%NaCl solution.And HPS1-D proved to be a neutral sugar, with→4) α-D-Glcp (l→,→4,6) α-D-Glcp (1→in backbone,and α-L-Araf (1→,→5) α-L-Araf (1→,→3,5) α-L-Araf (1→n branches. HPSl-D has a random coil state conformation with monodisperse mass distribution in0.9%NaCl solution.We used the classical pathway hemolysis to study anti-complement activity of HPS1, HPS1-A, HPS1-B, HPS1-CandHPS1-D.The results showed that HPS1-D, HPS1-A, HPS1-B anti-complement activity better, HPS1followed, HPS1-C activity is weakest.更多还原