摘要
【目的】探讨白杨素和柚皮素抗猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)的作用靶点和机制,为进一步以白杨素和柚皮素开发新型抗PEDV的治疗药物提供理论依据。【方法】利用PharmMapper、TCMSP、SEA Search Server和STITCH在线数据库获得白杨素和柚皮素的潜在作用靶点,同时检索GeneCards数据库获得PEDV对宿主的作用靶点,利用在线程序Draw Venny Diagram获取白杨素、柚皮素与疾病的交集靶点。通过STRING数据库和Cytoscape 3.9.1软件构建靶蛋白互作(PPI)网络并筛选关键靶点,对靶点进行GO功能和KEGG通路富集分析。通过AutoDock Vina v 1.2.0软件对核心靶点及小分子进行分子对接,并分析白杨素和柚皮素与靶蛋白的结合能和结合模式,利用PyMOL v 2.5实现对接结果的可视化。采用实时荧光定量PCR和Western blotting检测白杨素和柚皮素对核心靶蛋白表达的影响。【结果】白杨素潜在抗PEDV的靶点有12个,其中白蛋白(ALB)、雌激素受体1(ESR1)、转化生长因子β-1蛋白(TGF-β1)可能是白杨素抗PEDV的核心靶点;柚皮素潜在抗PEDV的靶点有18个,其中ALB、胱天蛋白酶3(Caspase-3,CASP3)、过氧化物酶体增殖物激活受体γ(PPARG)可能是柚皮素抗PEDV的核心靶点。白杨素抗PEDV靶点涉及21种生物过程、5种细胞组分和6种分子功能,共获得11条信号通路;柚皮素抗PEDV靶点涉及31种生物过程、8种细胞组分和19种分子功能,共获得13条信号通路。核心靶点与两种天然化合物之间以氢键和疏水作用力为主,有较强的相互作用。白杨素和柚皮素能极显著降低Caspase-3表达量(P<0.01),可能通过影响Caspase-3的表达和活化来颉颃PEDV诱导的细胞凋亡;极显著升高TGF-β1蛋白表达量(P<0.01),可能通过影响TGF-β1的表达抗PEDV诱导的炎症反应而治疗PEDV的感染。【结论】本研究揭示了白杨素和柚皮素分别通过潜在核心靶点ALB、ESR1、TGF-β1和ALB、Caspase-3�
【Objective】This study was objective to explore the target and mechanism of chrysin and naringenin against Porcine epidemic diarrhea virus(PEDV),and provide theoretical basis for further developing new anti-PEDV therapeutic drugs with chrysin and naringenin.【Method】Online databases of PharmMapper,TCMSP,SEA Search Server and STITCH were used to obtain the potential targets of chrysin and naringenin,at the same time GeneCards database was searched to obtain the targets of PEDV on the host,and Draw Venny Diagram online program was used to obtain the intersection targets of chrysin,naringenin and disease.The target protein-protein interaction(PPI)network was constructed by STRING database and Cytoscape 3.9.1 software,and the key targets were screened,and the GO function and KEGG pathway enrichment of the targets were analyzed.The molecular docking of core targets and small molecules was carried out by AutoDock Vina v 1.2.0 software,and the binding energy and binding mode of chrysin and naringenin with target proteins were analyzed,and the docking results were visualized by PyMOL v 2.5.The effects of chrysin and naringenin on the expression of core target proteins were detected by Real-time quantitative PCR.【Result】The results showed that there were 12 potential anti-PEDV targets of chrysin,among which albumin(ALB),estrogen receptor 1(ESR1)and transforming growth factorβ-1 protein(TGF-β1)might be the core targets of chrysin against PEDV.Naringenin had 18 potential anti-PEDV targets,among which ALB,Caspase-3(CASP3)and peroxisome proliferator-activated receptorγ(PPARG)might be the core targets of naringenin against PEDV.The anti-PEDV target of chrysin involved 21 biological processes,5 cellular components and 6 molecular functions and obtained 11 signal pathways,while naringenin anti-PEDV targets involved 31 biological processes,8 cellular components and 19 molecular functions,and obtained 13 signal pathways.There was a strong interaction between the core targets and the two natural compounds due to hydrog
作者
植玉鹏
刘雨桐
陈弟诗
宫萌菲
夏学妹
任玉鹏
ZHI Yupeng;LIU Yutong;CHEN Dishi;GONG Mengfei;XIA Xuemei;REN Yupeng(College of Animal Science and Veterinary Medicine,Southwest Minzu University,Chengdu 610041,China;Sichuan Provincial Center for Animal Disease Prevention and Control,Chengdu 610041,China)
出处
《中国畜牧兽医》
CAS
CSCD
北大核心
2024年第4期1757-1772,共16页
China Animal Husbandry & Veterinary Medicine
基金
研究生创新型科研项目(YB2023352)
四川省科技计划项目重点研发项目(2023YFN0021)。
