Li Ke, Cao Y-X, Jiao S-M, Du G-H, Du Y-G, Qin X-M
Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan, China.
Institute of Process Engineering, Chinese Academy of Sciences, Beijing, China.
Front Pharmacol. 2020 Nov 24;11:582091. doi: 10.3389/fphar.2020.582091. eCollection 2020.
Saccharides are the most abundant substance with the strongest immunological activity in Astragali Radix (AR). However, systematic structure study and immunoactivity screening of polysaccharides with different molecular weights (Mw) in AR have yet to be conducted. In this study, Astragalus polysaccharides (APSs) were divided into three fragments of different Mw values, >2,000 kDa (APS-Ⅰ), about 10 kDa (APS-Ⅱ), and about 300 Da (APS-Ⅲ), by using ultrafiltration for the first time. The structural differences of the three products were determined on the basis of monosaccharide composition, FT-IR spectrum, linkage analysis, and nuclear magnetic resonance analysis. Cellular immune activity experiments and cyclophosphamide immunosuppression animal model experiments for nonspecific and specific immunoactivity screening were applied to identify the most immunogenic fragment in APSs. Linkage analysis results showed that APS-Ⅰ, APS-Ⅱ, and APS-Ⅲ have different attachment sites of monosaccharide residues. Immune screening experiments indicated that the Mw of the APSs influenced their activity, and APS-Ⅱ had the strongest immunoenhancing activity among the products. This research may serve as a reference for further study on APSs with different structures and immune activities, and as a guidance for the quality control of APSs and the development of new APS products.
糖类是黄芪中含量最为丰富且免疫活性最强的物质。然而,尚未对黄芪中不同分子量(Mw)的多糖进行系统的结构研究和免疫活性筛选。在本研究中,首次采用超滤法将黄芪多糖(APSs)分为分子量不同的三个片段,即>2000 kDa(APS-Ⅰ)、约10 kDa(APS-Ⅱ)和约300 Da(APS-Ⅲ)。基于单糖组成、傅里叶变换红外光谱(FT-IR)、连接分析和核磁共振分析确定了这三种产物的结构差异。应用细胞免疫活性实验和环磷酰胺免疫抑制动物模型实验进行非特异性和特异性免疫活性筛选,以鉴定APSs中免疫原性最强的片段。连接分析结果表明,APS-Ⅰ、APS-Ⅱ和APS-Ⅲ的单糖残基连接位点不同。免疫筛选实验表明,APSs的分子量影响其活性,且APS-Ⅱ在这些产物中具有最强的免疫增强活性。本研究可为进一步研究具有不同结构和免疫活性的APSs提供参考,并为APSs的质量控制和新型APS产品的开发提供指导。