灰树花水溶物对EV71病毒的抑制作用  

Inhibitory effect of Grifola frondosa water soluble extracts on EV71 virus

作　　者：林震山[1,2] 熊雯宇 何君强 江小琴 李泉岑[1] 刘斌 LIN Zhenshan;XIONG Wenyu;HE Junqiang;JIANG Xiaoqin;LI Quancen;LIU Bin(College of Food Science,Fujian Agriculture and Forestry University;National Engineering Research Center of JUNCAO Technology,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学食品科学学院 [2]国家菌草工程技术研究中心,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2024年第3期410-418,共9页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建农林大学营养与功能性食品服务团队项目(11899170148);福建省科学技术厅区域发展项目(2020N3002);福建省科学技术厅对外合作项目(2021I0008);福建省重大专项(2021NZ0101)。

摘　　要：【目的】探讨灰树花水溶物(GFWE)的化学成分及其对EV71病毒的抑制作用,为促进灰树花综合利用、提高其经济附加值,以及开发预防和治疗EV71病毒感染的功能性食品提供依据。【方法】采用色谱法测定了GFWE的主要化学成分组成、多糖组成和分子质量。基于体外细胞培养采用CCK-8法确定了GFWE的安全上样量;以EV71病毒感染Vero细胞为模型,探究GFWE对EV71病毒的抑制率及对Vero细胞形态的影响,并采用荧光定量PCR检测了EV71病毒衣壳蛋白(VP1)编码基因mRNA的表达水平,初步评价了GFWE对EV71病毒的抑制作用。【结果】GFWE主要由低分子质量多糖、氨基酸、肽及其衍生物组成。GFWE富含的13种氨基酸中包含7种必需氨基酸;其多糖由葡萄糖、半乳糖、盐酸氨基葡萄糖和古罗糖醛酸4种单糖组成,物质的量比为0.907∶0.038∶0.029∶0.026,重均分子质量分别为15.67、10.10、6.60 ku和<1.00 ku。GFWE表现出较高的抗EV71病毒活性,100、200μg·mL^(-1) GFWE对EV71病毒的抑制率分别为91.32%、91.83%,25~400μg·mL^(-1) GFWE能显著降低EV71 VP1 mRNA的表达水平。【结论】GFWE主要由低分子质量多糖、氨基酸、肽及其衍生物组成,具有较高的抗EV71病毒活性,能够抑制感染EV71病毒的Vero细胞中VP1的合成。【Objective】The chemical composition of water soluble extracts of Grifola frondosa(GFWE) and its inhibitory effect on EV71 virus were evaluated to provide theoretical foundation for the comprehensive utilization of G.frondosa,improvement of its economic value added,and development of functional food featured on the prevention and treatment of EV71 virus.【Method】The main chemical composition,polysaccharide composition and molecular weight of GFWE were determined by chromatography,and the safe loading content of GFWE was determined via in vitro cell culture by CCK-8 method.Then the inhibition rate of GFWE on EV71 virus and its impact on cell morphology were determined using EV71 virus-infected Vero cells as the model;meanwhile,the expression level of EV71 VP1 mRNA was detected by fluorescence quantitative PCR to preliminarily assess the inhibitory effect of GFWE on EV71 virus.【Result】GFWE was mainly composed of low molecular weight polysaccharides,amino acids,peptides and their derivatives,and the 13 types of amino acids it contained included 7 essential amino acids.The polysaccharides were composed of glucose,galactose,glucosamine hydrochloride and guluronic acid,with a molar ratio of 0.907∶0.038∶0.029∶0.026,and average molecular weights of 15.67,10.10,6.60 and <1.00 ku,respectively.GFWE showed relatively high inhibitory activity towards EV71 virus,with the inhibitory rates on EV71 virus reaching 91.32% and 91.83% at the GFWE concentrations of 100 and 200 μg·mL^(-1),respectively;meanwhile,the mRNA expression levels of VP1 were declined significantly at the GFWE concentrations of 25-400 μg·mL^(-1).【Conclusion】GFWE was mainly composed of low molecular weight polysaccharides,amino acids,peptides and their derivatives.It had high inhibitory activity on EV71 virus,and can reduce the synthesis of VP1 in EV71 virus-infected cells.

关 键 词：灰树花水溶物 多糖组成 蛋白质组成 抗EV71病毒活性 

分 类 号：TS219[轻工技术与工程—粮食、油脂及植物蛋白工程]