黄芪活性多糖APS-Ⅱ酶解方法的建立及其降解寡糖的免疫活性研究  

Establishment of enzymatic hydrolysis method of active Astragalus polysaccharides APS-Ⅱand study on the immune activity of oligosaccharides after its degradation

作　　者：李科[1,2,3,4] 崔连杰 李晓霞 石丽霞[1,3,4] 李震宇 秦雪梅[1,3,4] 杜昱光 LI Ke;CUI Lian-jie;LI Xiao-xia;SHI Li-xia;LI Zhen-yu;QIN Xue-mei;DU Yu-guang(Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan 030006,China;Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,China;Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education,Shanxi University,Taiyuan 030006,China;Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province,Shanxi University,Taiyuan 030006,China;College of Chemistry and Chemical Engineering,Shanxi University,Taiyuan 030006,China;Shanxi Fruit Industry Work Station,Taiyuan 030001,China)

机构地区：[1]山西大学中医药现代研究中心,山西太原030006 [2]中国科学院过程工程研究所,北京100190 [3]山西大学化学生物学与分子工程教育部重点实验室,山西太原030006 [4]地产中药功效物质研究与利用山西省重点实验室,山西太原030006 [5]山西大学化学化工学院,山西太原030006 [6]山西省果业工作总站,山西太原030001

出　　处：《药学学报》2021年第7期1936-1944,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81872962);国家博士后科学基金资助项目(2019M650851);山西省优秀人才科技创新项目(201605D211030,201705D211020);国家重点研发计划(2019YFC1710800).

摘　　要：黄芪多糖是黄芪中免疫调节活性主要物质,由于多糖结构难于精确测定,限制了黄芪多糖体内分子作用机制的深入研究。“多糖受体学说”认为免疫活性多糖分子中存在一种或多种寡糖片段“活性中心”,因此将黄芪多糖降解为寡糖,从寡糖水平研究多糖的活性中心,为黄芪多糖结构及作用机制研究提供了新思路。本文采用内切α-1,4-葡聚糖酶酶解,通过单因素试验和正交试验确定最佳降解条件,将本课题组已制备的免疫活性多糖APS-Ⅱ(10 kDa组分)降解为不同聚合度寡糖组分,通过聚丙烯酰胺凝胶色谱制备及特异性和非特异性免疫细胞实验,对不同寡糖组分进行免疫活性筛选。动物福利和实验过程均遵循山西大学动物伦理委员会的规定。结果发现,与免疫活性多糖APS-Ⅱ相比,不同寡糖组分在不同免疫活性上有明显差异。本文通过寡糖水平研究黄芪多糖的不同免疫活性,为进一步阐明黄芪多糖结构与功能奠定了基础,丰富了多糖受体学说,也为黄芪多糖的开发提供了新思路。Astragalus polysaccharides are the main immunomodulatory substances in Astragali Radix.The structure of polysaccharides is difficult to accurately determine,which limits the in-depth study of the molecular mechanism of Astragalus polysaccharides in vivo."Polysaccharide receptor theory"believes that there are one or more oligosaccharide fragment"active centers"in immunologically active polysaccharide molecules.Therefore,the degradation of Astragalus polysaccharides into oligosaccharides and the study of the active centers of polysaccharides at the oligosaccharide level provide new ideas in the study of the structure and mechanism of Astragalus polysaccharides.This article adopts endo-α-1,4-glucanase enzymatic hydrolysis,and determines the best degradation conditions through single factor test and orthogonal test to degrade the immunologically active polysaccharide APS-Ⅱ(10 kDa component)into oligomers with different degrees of polymerization.Then through the preparation of polyacrylamide gel chromatography and specific immune and non-specific immune cell tests,the immune activity screening of different oligosaccharide components is carried out.The animal welfare and the experimental process in this study follow the requirements of the Animal Ethics Committee of Shanxi University.The results showed that compared with the immunologically active polysaccharide APS-Ⅱ,different oligosaccharide components have obvious differences in different immunological activities.This paper studies the different immunological activities of Astragalus polysaccharides at the level of oligosaccharides,laying a foundation for further elucidating the structure and function of Astragalus polysaccharides,enriching the theory of polysaccharide receptors,and providing new ideas for the development of Astragalus polysaccharides.

关 键 词：黄芪多糖 酶解 免疫活性 寡糖 多糖受体学说 

分 类 号：R967[医药卫生—药理学]