淫羊藿多糖组成分析及其免疫调节作用研究

Composition and immunomodulatory activity of neutral and acidic polysaccharides isolated from Epimedii Folium

淫羊藿具有免疫调节、抗肿瘤、抗氧化等多种药理活性。多糖是淫羊藿物质基础的主要组分之一,且多糖的活性与结构密切相关。该研究通过DEAE-52纤维素柱及Sephadex G-100分离纯化得到淫羊藿中性多糖EPS-1-1和淫羊藿酸性多糖EPS-2-1。采用高效凝胶色谱法、完全酸水解、FT-IR分析、SEM检测以及刚果红实验等方法对2种多糖进行结构解析,并通过测定淫羊藿多糖对骨髓来源的小鼠树突状细胞(BMDC)成熟以及炎性细胞因子释放的影响,对其体外免疫调节活性进行研究。结果表明,EPS-1-1由果糖、甘露糖、核糖、鼠李糖、葡萄糖、半乳糖、木糖、阿拉伯糖组成,物质的量比为1.90∶0.67∶0.05∶0.08∶3.29∶1.51∶0.05∶0.37。EPS-2-1由物质的量比为5.25∶0.18∶0.32∶0.13∶1.14∶0.16∶0.55∶0.08∶0.2的果糖、甘露糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、木糖和阿拉伯糖组成。体外研究结果表明,EPS-1-1和EPS-2-1可以通过上调骨髓来源的小鼠树突状细胞中MHCⅡ和共刺激因子(CD40、CD80、CD86)的表达,提高炎性细胞因子(IL-6、TNF-α、IL-12)的水平,促进骨髓来源的小鼠树突状细胞的成熟和功能,提示淫羊藿多糖EPS-1-1和EPS-2-1均具有很好的免疫调节活性。

Epimedii Folium possesses many pharmacological activities including immunomodulation, anti-oxidation, and anti-tumor. Polysaccharides are the main components of Epimedii Folium, and their activities are closely related to the structure. The present study isolated a neutral polysaccharide(EPS-1-1) and an acidic polysaccharide(EPS-2-1) from the aqueous extract of Epimedii Folium through DEAE-52 cellulose anion-exchange chromatography and Sephadex G-100. The structures were characterized by chemical composition analysis, high-performance gel permeation chromatography(HPGPC), Fourier-transform infrared spectrometry(FT-IR), 1-phenyl-3-methyl-5-pyrazolone(PMP) derivatization, scanning electron microscopy(SEM), Congo red test, etc. The immunomodulatory activity of polysaccharides in vitro was determined by investigating the effects on the maturation of bone marrow-derived dendritic cells(BMDCs) and the release of inflammatory cytokines. According to the structural characterization analysis, EPS-1-1 was composed of fructose(Fuc), mannose(Man), ribose(Rib), rhamnose(Rha), glucose(Glc), galactose(Gal), xylose(Xyl), and arabinose(Ara) at 1.90∶0.67∶0.05∶0.08∶3.29∶1.51∶0.05∶0.37(molar ratio), while EPS-2-1 was mainly composed of Fuc, Man, Rha, glucuronic acid(GlcA), galacturonic acid(GalA), Glc, Gal, Xyl, and Ara at 5.25∶0.18∶0.32∶0.13∶1.14∶0.16∶0.55∶0.08∶0.2. EPS-1-1 and EPS-2-1 could promote the maturation and function of BMDCs through up-regulating the expression of MHC-Ⅱ, CD86, CD80, and CD40, and increasing the levels of inflammatory cytokines(IL-6, IL-12, and TNF-α) in vitro experiments, which suggested that EPS-1-1 and EPS-2-1 possessed good immunomodulatory activity.