灰树花胞外多糖的结构及免疫调节活性  

Structure and immunomodulatory activity of extracellular polysaccharide from Grifola frondosa

作　　者：韩丽荣[1] 程代[1] 王莉蕊 王春玲[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院,天津300457

出　　处：《生物工程学报》2016年第5期648-656,共9页Chinese Journal of Biotechnology

基　　金：国家科技支撑计划(No.2012BAD33B04);天津应用基础重点项目(No.13JCZDJC29800)资助~~

摘　　要：对灰树花胞外多糖A组分(EXGFP-A)进行结构分析和免疫活性的研究。结构分析结果表明,EXGFP-A是一种主要含有葡萄糖的吡喃型中性多糖。气相结果说明EXGFP-A的单糖组成为鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖,摩尔比为0.28∶0.31∶0.30∶0.06∶7.98∶0.61。MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo-lium bromide)比色实验表明,当EXGFP-A浓度为80μg/m L,作用48 h时,RAW264.7细胞增殖指数达到最大值,为137.5%。吖啶橙(AO)染色结果表明EXGFP-A能够激活RAW264.7细胞,增强细胞内部核酸代谢水平。EXGFP-A在一定浓度范围内,可以提高RAW264.7细胞中NO的释放量,上调细胞内TNF-α、IL-1β、IL-6、IL-12、IFN-γ细胞因子以及细胞中iNOS的mRNA水平的表达。结果表明,EXGFP-A具有一定的免疫调节活性,为灰树花胞外多糖的结构分析和应用提供了科学依据。We aimed at analyzing the structure of extracellular polysaccharide A from Grifola frondosa（EXGFP-A） and testing its immunomodulatory activity. Structural analysis shows that EXGFP-A was a contained α-D-glucoside bond and pyranose ring. GC analysis reveals that EXGFP-A was mainly composed of rhamnose, arabinose, xylose, mannose, glucose, galactose, by the molar ratio of 0.28：0.31：0.30：0.06：7.98：0.61. The results of MTT（3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide） assay indicates when EXGFP-A was at a concentration of 80 μg/m L and treatment time of 48 h, RAW264.7 cells proliferation index reached a maximum of 137.5%. Meanwhile, the AO staining showed that EXGFP-A activated RAW264.7 cells and improved the level of intracellular nucleic acid metabolism. In addition, in a certain range of concentration, EXGFP-A was able to increase the release of NO in RAW264.7 cells, and upregulate the m RNA expression of immunological factor TNF-α, IL-1β, IL-6, IL-12, IFN-γ and i NOS of RAW264.7 cells. Our results confirm that EXGFP-A had immunomodulatory activity. Our findings provided scientific basis for the structural analysis and application of Grifola frondosa polysaccharide.

关 键 词：灰树花多糖 结构分析 RAW264.7细胞 免疫活性 

分 类 号：Q946[生物学—植物学]