摘要
从金针菇子实体中分离纯化多糖,并对多糖结构和体外免疫活性进行研究。采用水提醇沉法从金针菇子实体中提取粗多糖,利用DEAE‐Cellulose‐52及Sephacryl S‐300HR柱层析纯化得到FVPⅠ‐a,再利用HPLC‐ELSD技术、红外及核磁共振对FVPⅠ‐a进行结构解析。在体外以促RAW264.7巨噬细胞产NO、分泌细胞因子,促进小鼠淋巴细胞增殖实验,考察FVPⅠ‐a增强免疫的能力。从金针菇子实体中分离纯化得到FVPⅠ‐a,其为分子量81.4kDa,由葡萄糖、果糖和鼠李糖组成的β构型的吡喃型杂多糖。体外免疫实验表明,FVPⅠ‐a能够促进RAW264.7巨噬细胞产生NO及分泌细胞因子(IL‐1β,IL‐6,TNF‐α),能单独的促进小鼠淋巴细胞增殖(P<0.05),并能协同增强ConA和LPS对小鼠淋巴细胞的促增殖作用(P<0.01,P<0.05)。首次从金针菇子实体中获得FVPⅠ‐a杂多糖,其在体外具有增强非特异性免疫反应及增强特异性免疫反应的能力。
The polysaccharides from the fruiting body of Flammulina velutipes were extracted, and the immunity-enhancing activities of the polysaccharides in vitro were evaluated. By water extraction and alcohol precipitation, crude polysaccharides were obtained. Through purification by DEAE-Cellulose 52 ion exchange chromatography and SephacrylS-300 HR gel filtration chromatography, a polysaccharide named as FVP I-a was obtained. HPLC-ELSD technology, IR and NMR were used to evaluate the structure of FVP I-a. NO production of RAW264.7 macrophages, cytokines secretion and lymphocyte proliferation were used as the indicator to evaluate the immunity-enhancing activities of F. velutipes polysaccharides in vitro. FVP I-a was mainly composed of glucose, a trace amount of fructose and rhamnose, and the molecular weight was 81.4kDa. IR analysis indicated that FVP I-a was a β-pyranoside polysaccharides, and NMR analysis also hinted that FVP I-a was a β-polysaccharides. FVP I-a can largely promote the NO production and augment the IL-1β, IL-6, TNF-α secretion of RAW264.7 macrophages. FVP I-a can promote lymphocyte proliferation (P〈0.05), and synergistically enhance the effect of ConA and LPS on the proliferation of mouse lymphocytes (P〈0.01, P 〈0.05). The FVP Ⅰ-a heteropolysaccharide was obtained from F. velutipes for the first time, and it could enhance non-specific immune response and specific immune responses in vitro.
出处
《菌物学报》
CAS
CSCD
北大核心
2014年第2期375-384,共10页
Mycosystema
基金
四川省精深加工研究岗位建设项目[川农业(2009)75号]
菌类药材研究与开发四川省青年科技创新团队(No.2011JTD0021)
四川省重大科技计划项目食药用菌现代产业链关键技术研究集成与产业化示范
农业微生物平台项目
四川省十二五育种攻关项目菌类药材优质种质资源的收集及新材料的选育(No.2011NZ0098‐12‐08)
关键词
金针菇
杂多糖
结构
一氧化氮
细胞因子
免疫
Flammulina velutipes, heteropolysaccharides, structure, nitric oxide, cytokines, immune