网格状与线状鼠尾藻多糖抗炎作用研究

Anti-inflammation effects of linear and mesh-shaped polysaccharides from Sargassum thunbergii

目的对鼠尾藻多糖进行分离纯化,并对其理化特性、微观结构和对炎症因子表达的影响进行研究。方法采用热水提取法及DEAE-Sepharose CL-6B和Sephacryl S-300HR柱层析分离纯化鼠尾藻多糖,利用高效空间排阻色谱法(HPSEC)、气相色谱法(GC)、红外光谱法(FTIR)和透射电子显微镜(TEM)研究理化特性及微观结构,实时荧光定量PCR法研究炎症相关因子的mRMA表达。结果得到两种鼠尾藻均一多糖STP-Ⅰ和STP-Ⅲ,相对分子质量分别是29和350kDa,糖含量均超过96%。透射电子显微镜观察到STP-Ⅰ是网格状结构,STP-Ⅲ是线状且螺旋缠绕结构。活性研究表明,STP-Ⅲ能更好地抑制脂多糖(LPS)诱导下小鼠巨噬细胞RAW264.7系炎症因子TNF-α、IL-6和COX-2的mRNA表达量,在150μg·mL^(-1)质量浓度下,STPⅢ对炎症因子TNF-α、IL-6和COX-2的mRNA表达抑制率均超过95%。结论鼠尾藻多糖具有抗炎症因子表达的作用,可以作为潜在的抗炎药物开发。线状缠绕的鼠尾藻多糖STP-Ⅲ较网格状的STP-Ⅰ能更好地抑制炎症相关因子的表达。

Objective To isolate polysaccharides fromSargassum thunbergii and study their physicochemical properties,microstructures and anti-inflammation effects.Methods Two polysaccharides(STP-I and STP-Ⅲ)were purified using water extraction method,DEAE-Sepharose CL-6 Band Sephacryl S-300 HR column chromatogram.The physicochemical properties and microstructures were studied using high-performance size-exclusion chromatography(HPSEC),gas chromatography(GC),Fourier transform infrared spectroscopy(FTIR)and transmission electron microscopy(TEM)analysis.Anti-inflammation activities of the polysaccharides were identified using real time quantitative PCR.Results STP-Ⅰand STP-Ⅲwere all homogeneous,molecular weights(Mw)were 29 and 350 kDa,and the contents of total carbohydrates were more than 96%.TEM revealed further that STP-Ⅰwas mesh-shaped,and STP-Ⅲwas observed as linear microstructures with entangled chains.Due to the results of anti-inflammation activities of the polysaccharides,STP-Ⅲshowed the higher inhibitory effects of TNF-α,IL-6 and COX-2 mRNA expressions in LPS-stimulated RAW 264.7 mouse macrophage cells than others,and the inhibitory effects were more than 95%at 150μg·mL-1 concentrations.Conclusion The polysaccharides fromS.thunbergii have obvious anti-inflammation effects and may serve as potential anti-inflammatory agents,and linear STP-Ⅲshows more noticeable anti-inflammation effects than the huge meshshaped STP-Ⅰ.