猴头菇多糖对MDRV感染RAW264.7细胞后病毒复制及TLR3诱导表达的影响  

Effect of Hericium erinaceus polysaccharide on viral replication and TLR3 induced expression of MDRV-infected RAW264.7 cells

作　　者：林绍青 骆钰 李明慧 孙雪宁[1] 吴宝成[1,2] 黄一帆 吴异健[1,2] LIN Shao-qing;LUO Yu;LI Ming-hui;SUN Xue-ning;WU Bao-cheng;HUANG Yi-fan;WU Yi-jian(College of Animol Science , Fujian Agriculture and Forestry University, Fuzhou 350002, China;Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fuzhou 350002, China)

机构地区：[1]福建农林大学动物科学学院,福建福州350002 [2]福建省兽医中药与动物保健重点实验室,福建福州350002

出　　处：《中国兽医科学》2019年第6期773-780,共8页Chinese Veterinary Science

基　　金：福建省自然科学基金项目(2017J01597)

摘　　要：本试验旨在探索猴头菇多糖(HEP)对番鸭呼肠孤病毒(MDRV)感染RAW264.7细胞中病毒复制及TLR3诱导表达的影响,为进一步解析HEP在调节MDRV所致的番鸭机体免疫抑制中的作用机制提供数据参考。本试验首先通过测定HEP在小鼠单核巨噬细胞白血病细胞(RAW264.7细胞)上的安全浓度的基础上确定HEP的添加浓度;然后通过RT-qPCR方法和Western-blot方法分别检测各组细胞中TLR3 m RNA的转录水平和TLR3、病毒σNS的蛋白表达量,从基因和蛋白的分子层面分析HEP对MDRV感染RAW264.7细胞的免疫调节作用。结果显示:HEP在46.88~187.5μg/m L浓度范围内是可以促进RAW264.7细胞生长,浓度为93.75μg/m L时细胞相对增殖率最高,VCG细胞TLR3 m RNA的相对表达量和蛋白表达量均极显著(P<0.01)高于BCG组,HPG与HTG细胞TLR3 m RNA的相对表达量和蛋白表达量均极显著(P<0.01)低于VCG组。结论:HEP可从基因与蛋白水平抑制MDRV感染早期过度活化的TLR3的表达,从而起到抗病毒作用。The aim of this study was to investigate the effect of Hericium erinaceus polysaccharide(HEP) on viral replication and TLR3-induced expression in RAW264.7 cells infected with Muscovy duck reovirus(MDRV),and to provide a data reference for further analysis of the mechanism of action of HEP in regulating the immune suppression of Muscovy ducks caused by MDRV.In this experiment wo first determined the concentration of HEP added by measuring the safe concentration of HEP on mouse mononuclear macrophage leukemia cells(RAW264.7 cells).Then,RT-q PCR and Western-blot were used to detect the transcription levels of TLR3 m RNA and the protein expression of TLR3 and σNS in each group.The immunomodulatory effects of HEP on MDRV-infected RAW264.7 cells were analyzed from the molecular level of genes and proteins.In result,HEP can promote the growth of RAW264.7 cells in the concentration range from46.88 μg/m L to 187.5 μg/m L.The relative proliferation rate of cells is the highest when the concentration is 93.75 μg/m L.The relative expression and protein expression of TLR3 m RNA in VCG cells were significantly higher(P<0.01) than those in the BCG group.The relative expression and protein expression of TLR3 m RNA in HPG and HTG cells were significantly(P<0.01) lower than those in the VCG group.In conclusion,HEP can inhibit the expression of over-activated TLR3 in the early stage of MDRV infection from the gene and protein levels,thus playing an antiviral role.

关 键 词：猴头菇多糖 番鸭呼肠孤病毒 RAW264.7细胞 TLR3受体 

分 类 号：S853.74[农业科学—临床兽医学]