摘要
【背景】灵芝是一种广受关注的药食两用真菌,在中国作为传统药材和健康食品已有几千年历史,近年来中国人工栽培灵芝规模扩展迅速,而多糖被认为是灵芝中最主要的活性成分。【目的】分离和纯化人工栽培灵芝子实体多糖,并对其结构和免疫调节活性进行研究。【方法】采用水提醇沉法提取灵芝子实体多糖(GLP),采用DE-52纤维素柱和Sephadex G-100进行多糖的分离纯化,高效凝胶渗透色谱法测定多糖分子量,1-苯基-3-甲基-5-吡啶啉酮(PMP)柱前衍生法测定单糖组成,红外光谱进行结构分析,噻唑蓝(MTT)比色法测定多糖对小鼠脾细胞增殖的影响,同时评价其对RAW 264.7细胞吞噬能力和细胞因子分泌能力的促进作用。【结果】GLP平均分子量为1.93×10~4 Da,单糖组成包含葡萄糖(Glc)、半乳糖(Gal)及甘露糖(Man),占比为Glc:Gal:Man=3.3:1.3:1.0。红外光谱显示,GLP的异头碳为β构型。GLP能直接促进脾细胞的增殖,且能显著增强刀豆蛋白A(Con A)诱导的T淋巴细胞和脂多糖(LPS)诱导的B淋巴细胞的增殖。此外,对于RAW 264.7细胞的吞噬能力及细胞因子分泌具有一定的促进作用。【结论】灵芝多糖可作为一个潜在的免疫调节药物而开发利用。
[Background] Ganodema lingzhi Sheng H. Wu, Y. Cao Y.C. Dai. is a widely regarded medicinal and edible fungus. Spectacularly, it was used as a traditional medicine and health food in China for thousands of years and the cultivation of G. lingzhi has been expanding rapidly recent years. Polysaccharides are considered to be the most important active ingredient in G. lingzhi. [Objective] This article focused on the extraction, purification, structure and immunoregulatory effects of a novel polysaccharide purified from the fruit bodies of G. lingzhi. [Methods] The polysaccharides of G.lingzhi（GLP） were extracted with boiling water and purified by DEAE cellulose-52 column and Sephadex G-100 column. The high performance gel permeation chromatography（HPGPC）, PMP derivatization and FT-IR method were used to analysis molecular weight, monosaccharide composition and backbone structure, respectively. The splenocyte proliferation was analysed by MTT assay. The potency of polysaccharides on the phagocytosis and cytokine secretion of RAW 264.7 cells was determined by commercial kit. [Results] The molecular weight of GLP was 1.93×10~4 Da, the monosaccharide composition of GLP was mainly composed of glucose, galactose and mannose, with a ratio of 3.3：1.3：1.0. The anomeric carbon of GLP has a β configuration. GLP resulted in a significant increase of lymphocyte proliferation and could also enhance the effect of Con A-induced T lymphocyte proliferation and the stimulation index of LPS-induced B lymphocytes. In addition, the phagocytosis and cytokine secretion of RAW 264.7 cells can be promoted by GLP. [Conclusion] GLP may be developed as a potential immunoregulatory drug.
作者
熊川
罗强
金鑫
陈诚
李强
陈祖琴
黄文丽
XIONG Chuan;LUO Qiang;JIN Xin;CHEN Cheng;LI Qiang;CHEN Zu-Qin;HUANG Wen-Li(Biotechnology and Nuclear Technology Research Institute, Sichuan Academy of Agricultural Sciences, Chengdu, Sichuan 610061, China;Institute for Viral Hepatitis, Key Laboratory of Molecular Biology for Infectious Diseases （Ministry of Education）, The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China;Institute of Plant Protection, Siehuan Academy of Agricultural Sciences, Chengdu, Sichuan 610066, China)
出处
《微生物学通报》
CAS
CSCD
北大核心
2018年第4期825-835,共11页
Microbiology China
基金
四川省科技支撑计划(2016GZ0355)~~
关键词
灵芝
多糖提取
分离纯化
免疫调节
Ganodema lingzhi, Extraction of polysaccharide, Isolation and purification,Immunoregulation
