太子参多糖的纯化、组成及对LPS损伤Raw264.7巨噬细胞的保护作用  

Purification and Composition of Polysaccharides from Pseudostellaria heterophylla and Protective Effect on Raw 264.7 Cells Induced by LPS

作　　者：谢美林 徐海清 李景明[1] 查道喜 王学瑛[2] 陈洋 朱启法 XIE Meilin;XU Haiqing;LI Jingming;ZHA Daoxi;WANG Xueying;CHEN Yang;ZHU Qifa(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China;Anhui Wannan Tobacco Leaf Co.,Ltd.,Xuancheng 242000,China)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083 [2]安徽皖南烟叶有限责任公司,安徽宣城242000

出　　处：《食品工业科技》2022年第14期392-400,共9页Science and Technology of Food Industry

基　　金：太子参脱毒示范种植及深加工研究(20190563004)。

摘　　要：太子参是我国传统中药材,为明确其多糖的组成与对Raw 264.7巨噬细胞的保护作用,本研究采用水提醇沉法提取其主要活性成分多糖,并通过阴离子交换柱层析纯化得太子参多糖(Pseudostellaria heterophylla polysaccharide,PHP);利用凝胶渗透色谱(GPC)法、高效液相色谱(HPLC)法测定其分子量、单糖组成,紫外光谱扫描、傅里叶红外光谱扫描初步解析其结构特征。进一步通过小鼠单核巨噬细胞白血病细胞(Raw 264.7)模型探究了PHP对LPS诱导损伤的保护作用。结果表明,太子参粗多糖的得率为8.23%,纯度为51.47%,而经DEAE-52柱层析分离纯化后得的PHP纯度达到了80.52%;PHP的分子量在500~92×10^(3) Da之间,单糖组成为葡萄糖、半乳糖、阿拉伯糖、甘露糖及葡萄糖醛酸。细胞实验确定80~640μg/mL为PHP的安全剂量,此浓度下可以提高脂多糖(Lipopolysaccharide,LPS)诱导损伤下的细胞存活率,640μg/mL的PHP剂量下存活率最大提升了7.21%,并且PHP可改善Raw 264.7巨噬细胞形态。综上,本研究提纯了太子参中多糖,对PHP进行了结构表征,并初步证实了PHP对LPS诱导损伤的Raw 264.7巨噬细胞有保护作用,这为太子参多糖的功能活性探索及构效、量效等深入研究奠定了基础。To determine the composition and protective effect on Raw 264.7 cells of polysaccharide from Pseudostellaria heterophylla as a traditional Chinese medicinal,the main active ingredient-Pseudostellaria heterophylla polysaccharide(PHP)was extracted by water,precipitated by ethanol,and purified by anion exchange column chromatography.Gel permeation chromatography(GPC)and high performance liquid chromatography(HPLC)were used to determine molecular weight and monosaccharide composition of PHP.Ultraviolet spectroscopy(UV)and Fourier transform infrared spectroscopy(FT-IR)were used for primary structural characterization of PHP.Furthermore,the Raw 264.7 cell model was used to explore protective effect of PHP on Raw 264.7 cells induced by LPS.The results showed that the yield of crude polysaccharide of Pseudostellaria heterophylla was 8.23%,and the polysaccharide content was 51.47%.After purification by DEAE-52 column chromatography,the polysaccharide content increased to 80.52%,and the molecular weight distribution was between 500 to 92×10^(3) Da.The monosaccharide components of PHP include glucose,galactose,arabinose,mannose and glucuronic acid.Cell experiments showed that the safe dose of PHP was 80~640μg/mL and could increase the survival rate of cells induced by lipopolysaccharide(LPS).At a dose of 640μg/mL,the survival rate increased by 7.21%,and it was found that PHP could improve the morphology of Raw 264.7 cells.In summary,in this study the polysaccharides from Pseudostellaria heterophylla were purified,the structure of PHP was characterized,and it was preliminarily confirmed that PHP had potential protective effect on LPS-induced damage to Raw 264.7 cells.This laid the foundation for the follow-up exploration of functional activity,structure-effect and dose-effect of PHP.

关 键 词：太子参多糖 单糖组成 Raw 264.7巨噬细胞 保护作用 

分 类 号：TS201.4[轻工技术与工程—食品科学]