板蓝根多糖对PRRSV-JXA1株体外增殖的影响  

Effects of Radix isatidis polysaccharides on the proliferation of PRRSV-JXA1 strain in vitro

作　　者：苏科 李倩楠 樊俊洋 刘健[1] 朱世豪 郝潇雅 杨明凡[1,2,3] 张红英 SU Ke;LI Qiannan;FAN Junyang;LIU Jian;ZHU Shihao;HAO Xiaoya;YANG Mingfan;ZHANG Hongying(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China;Henan Province Key Laboratory of Animal Food Safety,Zhengzhou 450046,China;Zhengzhou Key Laboratory of Research and Evaluation of Traditional Chinese Veterinary Medicine,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,河南郑州450046 [2]河南省动物性食品安全重点实验室,河南郑州450046 [3]郑州市中兽药创制与评价重点实验室,河南郑州450046

出　　处：《中国兽医学报》2022年第12期2505-2513,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30900472)。

摘　　要：为研究板蓝根多糖(Radix isatidis polysaccharide,IRPS)体外抑制猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)增殖的作用方式、复制环节和途径,通过荧光定量PCR进行了最佳计量和给药方式的筛选,确定了IRPS作用于病毒复制的环节,通过荧光定量PCR、间接免疫荧光、蛋白质印迹等试验探讨了TLR4/NF-ΚB途径对IRPS抑制PRRSV-JXA1株增殖的影响。结果显示,IRPS抑制PRRSV-JXA1株增殖的最佳剂量为0.015625 g/L,最佳给药方式和最佳阻断环节为预防作用和吸附环节,分别使病毒载量减少了91.34%(P<0.0001)和88.83%(P<0.0001);IRPS在多个时间点促进了PRRSV感染细胞TLR4表达;当TLR4/NF-ΚB途径被阻断时IRPS不能抑制PRRSV的增殖作用,反而促进其增殖(P<0.0001)。结果表明,IRPS主要作用于PRRSV-JXA1株感染的吸附环节,TLR4下游NF-ΚB信号通路是IRPS体外抑制PRRSV-JXA1株增殖的关键通路。This study aims to explore the antiviral mode and pathway of Radix isatidis polysaccharide(IRPS)against porcine reproductive and respiratory syndrome virus(PRRSV)in vitro.The optimal dose,mode of administration and key process of proliferation of PRRSV were screened and the role of TLR4/NF-κB pathway in the inhibition of IRPS on proliferation of PRRSV-JXA1 were investigated by quantitative fluorescent PCR,indirect immunofluorescence and Western blot.The results showed that the optimal dose of IRPS for inhibiting the proliferation of PRRSV-JXA1 was 0.015625 g/L,and the optimal administration mode was preventive and could block attachment,which reduced the viral load by 91.34%(P<0.0001)and 88.83%(P<0.0001)respectively.IRPS could promote TLR4 protein translation of cells infected with PRRSV at multiple time points when TLR4/NF-κB pathway was blocked,the inhibition of IRPS on PRRSV were vanished and even more promoted the proliferation of PRRSV-JXA1.In conclusion,IRPS inhibits the proliferation of PRRSV-JXA1 mainly by blocking virus attachment.The TLR4/NF-κB signaling pathway is the key pathway for IRPS to inhibit PRRSV-JXA1 proliferation in vitro.

关 键 词：板蓝根多糖 猪繁殖与呼吸综合征病毒 荧光定量PCR 间接免疫荧光 蛋白质印迹 TLR4 

分 类 号：S852.65[农业科学—基础兽医学] S859[农业科学—兽医学]