中国兽医科学2025，Vol.55Issue(4)：543-551,9.DOI:10.16656/j.issn.1673-4696.2025.0051
基于体外试验和网络药理学探究蓼大青叶抗PRRSV作用机制
Network pharmacology and in vitro experimental validation to reveal the mechanism of Polygoni Tinctorii Folium against PRRSV
摘要
Abstract
This study aimed to explore the effects,potential active ingredients and mechanism of Polygoni Tinctorii Folium(PTF)against porcine reproductive and respiratory syndrome virus(PRRSV)based on in vitro experiments and network pharmacology.Firstly,the water extract of the PTF was pre-pared and its cytotoxicity on Marc-145 cells was determined,and then the inhibitory effects of the PTF water extract on PRRSV proliferation at different times and doses were determined by RT-qPCR,West-ern-blot and indirect immunofluorescence assay.Secondly,the active ingredients and targets of the PTF were collected from the HERB and TCMSP databases,and the PRRSV diseases targets were collected from the CTD and GeneCards databases,then their intersection targets were obtained and the corresponding pro-tein-protein interaction network was constructed.Furthermore,GO function analysis and KEGG pathway enrichment analysis were conducted.Finally,the selected core targets and main active ingredients were verified by molecular docking.The results of in vitro experiments showed that the maximum cell safety concentration of the PTF water extract significantly inhibited the virus proliferation at 24,36 and 48 hours post PRRSV infection,and the low,medium and high concentrations of the PTF water extract could inhibit PRRSV proliferation in a dose-dependent manner.The results of network pharmacology showed that there are 10 active ingredients in the PTF,which can act on 46 PRRSV disease targets.GO function analysis yielded 148 entries,and KEGG pathway enrichment analysis yielded 116 signaling pathways.Molecular docking results showed that the main active ingredient of PTF,kaempferol,β-sitosterol and gomphrenol could bind well with all the core targets such as caspase 3(CASP3),prostaglandin endoperox-ide synthase 2(PTGS2),peroxisome proliferator-activated receptor γ(PPARG),estrogen receptor 1(ESR1),intercellular adhesion molecule 1(ICAM1)and transforming growth factor-β1(TGFB1),but β-sitosterol had no hydrogen bonding with CASP3,ESR1,ICAM1 and PPARG.This research results indicated that the PTF has direct anti-PRRSV effect and may exert indirect anti-PRRSV effect by acting on targets such as CASP3,PTGS2,PPARG,ESR1,ICAM1 and TGTB1 through active ingredients such as kaempferol and gomphrenol.关键词
蓼大青叶/猪繁殖与呼吸综合征病毒/网络药理学/分子对接Key words
Polygoni Tinctorii Folium/PRRSV/network pharmacology/molecular docking分类
农业科技引用本文复制引用
李佳明,张玲珑,黄鹤,刘雪威,李焕荣,董虹,周双海..基于体外试验和网络药理学探究蓼大青叶抗PRRSV作用机制[J].中国兽医科学,2025,55(4):543-551,9.基金项目
国家自然科学基金项目(32102663) （32102663）
家畜产业技术体系北京市家畜创新团队项目(BAIC05-2024) （BAIC05-2024）
北京市属高校分类发展-都市农林特色教师队伍建设项目(11000024T000002961733) （11000024T000002961733）
