Han Lirong, Cheng Dai, Wang Lirui, Wang Chunling
College of Food Engineering and Biological Technology, Tianjin University of Science & Technology, Tianjin 300457, China.
Sheng Wu Gong Cheng Xue Bao. 2016 May 25;32(5):648-656. doi: 10.13345/j.cjb.150399.
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/mL 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 mRNA expression of immunological factor TNF-α, IL-1β, IL-6, IL-12, IFN-γ and iNOS 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.
我们旨在分析灰树花胞外多糖A(EXGFP-A)的结构,并测试其免疫调节活性。结构分析表明,EXGFP-A含有α-D-糖苷键和吡喃糖环。气相色谱分析显示,EXGFP-A主要由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖组成,摩尔比为0.28:0.31:0.30:0.06:7.98:0.61。MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)检测结果表明,当EXGFP-A浓度为80μg/mL且处理时间为48小时时,RAW264.7细胞增殖指数达到最大值137.5%。同时,AO染色显示EXGFP-A激活了RAW264.7细胞并提高了细胞内核酸代谢水平。此外,在一定浓度范围内,EXGFP-A能够增加RAW264.7细胞中NO的释放,并上调RAW264.7细胞免疫因子TNF-α、IL-1β、IL-6、IL-12、IFN-γ和iNOS的mRNA表达。我们的结果证实EXGFP-A具有免疫调节活性。我们的研究结果为灰树花多糖的结构分析和应用提供了科学依据。