为研究灰树花发酵产物提取出的胞内多糖的免疫活性,采用50只雄性昆明小鼠,随机分为5组,每组10只,设100、200和400 mg/(kg·d)3个水平处理组,并设阳性对照组(黄芪多糖400 mg/(kg·d))和阴性对照组(生理盐水),饲养30 d后观察多糖对细胞免疫功能的影响。通过流式细胞仪检测脾细胞表面分子的CD4和CD8及对T淋巴细胞增殖的影响;用实时荧光定量PCR(RT-PCR)检测其细胞因子IL-6、IL-8、CRP和TNF-α的表达。结果表明:灰树花胞内多糖使小鼠脏器显著增大(P≤0.05);并显著增加了脾淋巴细胞的增殖能力(P≤0.05);对于T淋巴细胞亚群中免疫细胞CD4/CD8的比值随着用药量的增加而提升(P≤0.05);同时小鼠肠道中的细胞因子IL-6、IL-8和TNF-α的分子表达量起到了上调作用(P≤0.05);并且下调了炎症因子CRP的分子表达量(P≤0.05)。灰树花菌丝体胞内多糖能使小鼠体内免疫器官增大,免疫细胞分泌增多,相关免疫因子表达量增加。
To study the immune activity of extracted from fermentation products of Grifola frondosa,fifty male Kunming mice were randomly divided into five groups with ten mice in each.Three groups were treated with intracellular polysaccharide at 100,200 and 400 mg/(kg·d) levels respectively,a positive control group treated with Astragalus polysaccharides at 400 mg/(kg·d) and a negative control group was given normal saline.The effects of polysaccharide on cellular immunity function in mice were observed after 30 days.The effects of CD4 and CD8 on the proliferation of T lymphocytes were detected by flow cytometry.The expression of IL-6,IL-8,CRP and TNF-α was detected by real-time fluorescence quantitative PCR (RT-PCR).G. frondosa intracellular polysaccharides enhanced the proliferation of spleen lymphocyte (P≤0.05).The proportion of T lymphocyte subsets,CD4/CD8 in different treatments,was increased with the dose of intracellular polysaccharide.The expressions of IL-6,IL-8 and TNF-α were up-regulated in intestinal tract (P≤0.05),and the expression of CRP was decreased(P≤0.05).In conclusion,the intracellular polysaccharides from the mycelium of G. frondosa could enhance the immune organs in mice and increase the secretion of the immune cells and the expression of related factors.