绣球菌多糖表征及其抗氧化和免疫活性

提取纯化绣球菌多糖(Sparassis latifolia polysaccharides,SCPs),研究其表征和功能活性,探索绣球菌多糖表征与其抗氧化及免疫活性之间的关系。以绣球菌子实体为原料,采用聚能超声波辅助水提醇沉法提取绣球菌多糖,经DEAE-52、SephadexG-100纯化,用高效凝胶渗透色谱法、离子色谱法、傅里叶红外色谱法、扫描电镜、原子力显微镜对绣球菌多糖进行初步表征,检测绣球菌多糖清除DPPH、·OH、O2^-·自由基能力以及总还原力,用MTT法检测绣球菌多糖对巨噬细胞RAW264.7增殖的影响。结果表明,SCPs分子量范围为215Da–393kDa,由葡萄糖、甘露糖、半乳糖、木糖、果糖构成,摩尔比13:4:1:2:3,其表观形貌为簇状堆积,交织,结构规律性不强,表面光滑,呈一定的网络状结构,分子呈现链状构象,具有高度的分支结构,链间形成小环且伴随一定的球形颗粒。SCPs具有一定的还原能力和清除DPPH、·OH、O2^-·自由基的能力,且能够促进巨噬细胞RAW264.7的增殖。绣球菌多糖的抗氧化及免疫活性可能与其分子量、单糖组成、糖链分支及分子构象有关。

Characterization,antioxidant and immunity activities of Sparassis latifolia polysaccharides

This study aims at revealing characteristics and functional activity of Sparassis latifolia polysaccharides (SCPs), and exploring the relationship between the characteristics and antioxidant and immune activities of SCPs. SCPs were isolated and purified from Sparassis latifolia fruiting bodies by employing a procedure involving water extraction and alcohol precipitation assisted energy-gathered ultrasonic, DEAE-52 and Sephadex G-100. The structure of SCP was detected by using high performance gel permeation chromatography (HP-GPC), ion chromatography (IC), Fourier transformed infrared spectrometry (FT-IR), scanning electron microscope (SEM) and atomic force microscope (AFM). The antioxidant activities of SCPs were assayed through determining the DPPH, ·OH,$O_{2}^{-}$·free radical scavenging activity and total reducing power. The MTT assay was used to measure the effect of SCPs on proliferation of macrophages RAW264.7. The results showed that the molecular weight of SCPs was approximately 215Da-393kDa. SCPs were composed of glucose, mannose, galactose, xylose and fructose at the molar ratio of 13:4:1:2:3. The apparent morphology of SCPs was cluster accumulation, interweaving, irregular structure, smooth surface and a certain network structure. SCPs exhibited chain conformation with a highly branched structure, forming the small loops between chains with certain spherical particles. SCPs had certain reducing ability and showed effective DPPH, ·OH,$O_{2}^{-}$·free radical scavenging activities. SCPs could promote the proliferation of macrophages RAW264.7. The antioxidant and immune activities of SCPs might be related to their molecular weight, monosaccharide composition, carbohydrate branching and molecular conformation.