[关键词]
[摘要]
目的 对防风Saposhnikovia divaricata中分离得到均一多糖的结构及其免疫调节活性进行研究。方法 采用DEAE-纤维素、Sephadex G-75等柱色谱法,对防风多糖进行系统分离纯化,采用ESI-MSn、GC-MS、NMR等谱学技术,对分离得到的防风多糖SP800203的相对分子质量分布、单糖组成、寡糖片段、糖残基类型和糖苷键连接方式等进行分析,确定其结构。采用细胞实验和斑马鱼实验,研究防风多糖SP800203的免疫调节活性。结果 从防风中分离得到防风多糖SP800203,糖醛酸质量分数为75.73%,相对分子质量约为7.14×104,由鼠李糖、半乳糖、阿拉伯糖和半乳糖醛酸组成,单糖物质的量比为2.8∶6.7∶6∶84.5。主链由半乳糖醛酸聚合而成,2条支链分别由阿拉伯糖、半乳糖醛酸、半乳糖、鼠李糖,以及阿拉伯糖、半乳糖醛酸、半乳糖聚合而成,二者分别通过β、α糖苷键与主链相连。防风多糖SP800203可以促进巨噬细胞一氧化氮、肿瘤坏死因子-α、白介素-1β和白介素-6的释放,增加斑马鱼的免疫细胞密度和巨噬细胞数量。结论 防风多糖SP800203为从防风中分离得到的新的均一多糖,具有良好的免疫调节活性;研究结果为阐明防风免疫调节作用机制奠定了基础,为防风临床应用及进一步研究与开发提供科学依据。
[Key word]
[Abstract]
Objective To study the structure and immunomodulatory activity of homogeneous polysaccharides isolated from Fangfeng (Saposhnikoviae Radix).Methods Polysaccharide SP800203 was isolated from Saposhnikoviae Radix by several chromatographic techniques, such as DEAE-Cellulose and Sephadex G-75. The relative molecular weight, monosaccharide composition, oligosaccharide fragments, sugar residue types and glucosidic bond linkage modes were characterized with ESI-MSn, GC-MS, and NMR. Cell and zebrafish experiments were used to explore the immunomodulatory activity of polysaccharide SP800203. Results Polysaccharide SP800203 was isolated from Saposhnikoviae Radix, with a glucuronic acid content of 75.73%, a molecular weight of 7.14×104, composed of rhamnose (Rha), galactose (Gal), arabinose (Ara), and galacturonic acid (GalA), the molar ratio of monosaccharides was 2.8:6.7:6:84.5. The main chain was polymerized by GalA, and the two branch chains were separately polymerized by Ara, GalA, Gal, Rha and Ara, GalA, Gal, which were connected to the backbone by β and α glycosidic bonds, respectively. Polysaccharide SP800203 could significantly promote the release of NO, TNF-α, IL-1β, IL-6 and increase the density of immune cells and the number of macrophages. Conclusion Polysaccharide SP800203 is a new homogeneous polysaccharide isolated from Saposhnikoviae Radix, which shows good immunomodulatory activity. The results of the study lay a foundation for elucidating the mechanism of immune regulation of Saposhnikoviae Radix, and provide a scientific basis for the clinical application and further research of Saposhnikoviae Radix.
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[基金项目]
国家自然科学基金项目(82074283);中央高校基本科研业务费专项资金资助(2023-JYB-KYPT-15)