云芝葡聚糖对非洲猪瘟病毒的体外抑制作用  

In vitro antiviral effects of Coriolus versicolor-derived polysaccharides on African swine fever virus

作　　者：刘欣媛 孙元[1] 李素[1] 李连峰[1] 仇华吉[1] LIU Xinyuan;SUN Yuan;LI Su;LI Lianfeng;QIU Huaji(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150069

出　　处：《中国兽医学报》2022年第11期2133-2138,2144,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(32072855)。

摘　　要：研究云芝葡聚糖抑制非洲猪瘟病毒(ASFV)感染效果并初步探索其作用机制。利用CCK-8试剂检测云芝葡聚糖对原代猪肺泡巨噬细胞(PAM)的毒性并计算半数细胞毒性浓度(CC50);ASFV感染细胞,检测不同质量浓度云芝葡聚糖抗ASFV感染的剂量依赖效应,并计算半数抑制浓度(IC_(50));将25 mg/L的云芝葡聚糖与病毒悬液孵育处理,稀释后加入PAM,评价云芝葡聚糖体外直接杀灭ASFV的效果;同时在ASFV复制周期的不同时间点利用云芝葡聚糖处理细胞,确定其抗ASFV效果最显著的效应阶段;通过qPCR检测抗病毒相关细胞因子转录水平来初探其抗病毒机制。云芝葡聚糖的毒性较弱(CC50为270.1000 mg/L),具有一定的抗病毒效果(IC_(50)为0.9575 mg/L),其不能直接杀灭病毒;在ASFV感染前6~12 h使用云芝葡聚糖处理细胞,其抑制病毒复制的效果最为显著,且PAM的IL-1β、IL-6、IL-8及TNF-α等细胞因子的mRNA水平显著升高。This study aims to explore the inhibitory effects of Coriolus versicolor-derived polysaccharides(CVP)on African swine fever virus(ASFV)and its potential mechanism.The cytotoxicity of CVP was determined by CCK-8 assay and the 50%cell cytotoxicity(CC50)was calculated.Different concentrations of CVP were added to porcine alveolar macrophages(PAM)after ASFV infection to detect if it inhibits ASFV replication,and to calculate the 50%maximal inhibitory concentration(IC_(50))of CVP.CVP(25 mg/L)was incubated with ASFV suspension,and then the compound was added to PAM after dilution to determine if CVP has virucidal effects on ASFV in vitro.CVP(25 mg/L)was added to PAM at different time points of ASFV replication cycle to detect its most significant antiviral stage.The transcriptional levels of lots of cytokines were detected to tentatively explore the mechanism of CVP antiviral effects.The results showed that CVP had inhibitory effect on ASFV with a low cytotoxicity(CC50=270.1000 mg/L,IC_(50)=0.9575 mg/L),and it did not show direct virucidal effects on ASFV.In addition,CVP was able to activate inflammatory cytokines to exert anti-ASFV effects.

关 键 词：云芝葡聚糖 非洲猪瘟病毒 抗病毒药物 细胞因子 

分 类 号：S852.65[农业科学—基础兽医学]