中国畜牧兽医2024，Vol.51Issue(7)：3155-3165,11.DOI:10.16431/j.cnki.1671-7236.2024.07.039
天然产物库中体外颉颃A型塞内卡病毒化合物的筛选及作用途径研究
Screening and Action Pathway of Compounds Antagonistic Senacavirus A in vitro from Natural Product Library
摘要
Abstract
[Objective]The experiment was aimed to search for natural products with inhibitory activity against Senecavirus A(SVA)and further investigate its antagonistic pathway.[Method]The concentration of the natural products were uniformly diluted to 10 μmol/L.Combined with the recombinant Senecavirus A with luciferase(rSVA-NLuc),compounds with luciferase inhibitory activity were screened from the natural products library,and their activity was further verified using Real-time quantitative RT-PCR.The maximum non-toxic concentration was determined by cytotoxicity testing using lactate dehydrogenase(LDH)method.Virus titer determination,Real-time quantitative RT-PCR,Western blotting,IFA and other techniques were used to study the effect and pathway of antagonism against SVA.[Result]Five compounds,monensin sodium salt,progesterone,hypophyllanthin,4-hydroxyderricin and 2-methoxyestrone,which could antagonize SVA in vitro,were screened from 136 natural products.Progesterone,a natural product that could antagonize SVA in vitro,was screened by Real-time quantitative RT-PCR and cytotoxicity detection.Compared with blank control group,the mRNA expression level of SVA was extremely significantly decreased in vitro(P＜0.01),and the maximum non-toxic concentration reached 50 μmol/L.Further study showed that progesterone could continuously inhibit the proliferation of SVA at 8,12,24 and 36 h after infection,and the virus titer was significantly decreased at 24 and 36 h after infection compared with control group(P＜50.05).Compared with control group,different concentrations of progesterone(10,20 and 50 μmol/L)extremely significantly decreased the adsorption level of SVA at the adsorption stage(P＜0.01).In the replication stage,the level of SVA VP3 and virus titer were extremely significantly decreased(P＜0.01).The proportion of released extracellular virus extremely significantly decreased(P＜0.01).But there was no significant effect on other approaches.These results indicated that progesterone could inhibit the adsorption,replication and release of SVA in vitro,but it had no obvious effect on its induction and assembly pathway.[Conclusion]In this study,progesterone,a natural product with low cytotoxicity and excellent antagonistic effect against SVA,was screened from the small molecule library of natural products.The product could inhibit the adsorption,replication and release of SVA,thus preventing the infection of SVA.This study provided a scientific reference for promoting the development of anti-SVA drugs and studying the infection bias of SVA.关键词
孕酮/A型塞内卡病毒(SVA)/天然产物/颉颃途径Key words
progesterone/Senecavirus A(SVA)/natural product/antagonistic pathways分类
农业科技引用本文复制引用
王西彤,魏荣,尼博,黄保续,沙洲,迟田英,崔进,郑辉,陈峰,曹振山,张慧,戈胜强..天然产物库中体外颉颃A型塞内卡病毒化合物的筛选及作用途径研究[J].中国畜牧兽医,2024,51(7):3155-3165,11.基金项目
"十四五"国家重点研发计划"口蹄疫病毒的分子流行病学与传播机制"(2021YFD1800300) （2021YFD1800300）
中国动物卫生与流行病学中心创新基金(DW2021009) （DW2021009）
