基于生物信息学探讨猪繁殖与呼吸综合征病毒感染特征及银花甘草汤防治机制

doi:10.16431/j.cnki.1671-7236.2023.10.034

摘要/Abstract

摘要： 【目的】基于生物信息学分析猪繁殖与呼吸综合征病毒(PRRSV)感染后的生物标志基因,并对银花甘草汤进行成分鉴定及网络药理学研究,探究银花甘草汤防治PRRSV的作用机制。【方法】通过高通量基因表达(GEO)数据库收集PRRSV感染不同组织或机体的转录组数据,利用GEO2R在线软件筛选差异表达基因,并通过STRING软件构建蛋白互作网络,使用CytoNAC软件筛选核心蛋白,利用GO功能与KEGG通路富集分析确定感染后核心蛋白信号通路及生物学功能变化;选用超高效液相色谱飞行时间串联质谱(UPLC-Q-TOF-MS)分析并鉴定银花甘草汤组成成分,结合SwissTargetPredication数据库进行活性成分筛选及作用靶点预测,进一步通过网络药理学预测银花甘草汤对PRRSV感染的防治机制。【结果】GEO数据库的5组基因芯片共鉴定出4 476个PRRSV感染后的差异表达基因,蛋白互作分析共筛选出164个核心蛋白。PRRSV感染后的主要基因特征变化为非常规核糖体蛋白S27a(RPS27A)、泛素A-52 残基核糖体蛋白融合产物1(UBA52)和RPS家族,生物学功能变化为生物合成工艺和核糖体结构等,信号转导途径包括自噬、铁死亡及凋亡。银花甘草汤共包含70个有效成分(异甘草素、刺芒柄花素等),作用于PRRSV感染的基因共189个,涉及凋亡、细胞衰老等信号通路,其抗PRRSV感染的核心靶点包括肿瘤抑制蛋白53(TP53)、细胞外信号调节激酶1(MAPK1)、肿瘤坏死因子(TNF)、MAPK14等。【结论】PRRSV感染可能引起细胞自噬、铁死亡及凋亡,银花甘草汤有效成分可能通过调节TP53、TNF、CASP3和STAT3等基因以及细胞周期、凋亡等信号通路抵抗PRRSV感染。

关键词: 猪繁殖与呼吸综合征病毒(PRRSV); GEO数据库; 银花甘草汤; UPLC-Q-TOF-MS

Abstract: 【Objective】 Based on bioinformatics analysis,the biomarker genes of Porcine reproductive and respiratory syndrome virus (PRRSV) infection were analyzed,and the composition identification and network pharmacology of the Yinhua Gancao decoction were conducted to explore the mechanism of the treatment of PRRSV by the Yinhua Gancao decoction.【Method】 Transcriptome data of PRRSV infection in different tissues or bodies were collected using high-throughput gene expression (GEO) database,differentially expressed genes were screened using GEO2R online software,protein interaction network was constructed using STRING software,and core proteins were screened using CytoNAC software.GO function and KEGG pathway enrichment analysis were used to determine the changes of core protein signaling pathways and biological functions after infection.Ultra high performance liquid chromatography time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was used to identify the components of the Yinhua Gancao decoction,and the active ingredient screening and target prediction were carried out by the SwissTargetPredication database,and the prevention and control mechanism of the Yinhua Gancao decoction for PRRSV infection was further predicted by network pharmacology.【Result】 A total of 4 476 differentially expressed genes after PRRSV infection were identified by 5 groups of gene chips in GEO database,and 164 core proteins were screened by protein interaction analysis.After PRRSV infection,the main genetic characteristics of the non-conventional ribosomal protein S27a (RPS27A),ubiquitin A-52 residue ribosomal protein fusion product 1 (UBA52) and RPS family were changed,and the biological function changes were biosynthesis process and ribosome structure,etc.The signal transduction pathways included autophagy,iron death and apoptosis.The Yinhua Gancao decoction contained a total of 70 effective components (isoliquiritigenin,formononetin,etc.),which act on 189 genes of PRRSV infection,involving apoptosis,cell senescence and other signaling pathways.Its core targets against PRRSV infection included tumor suppressor protein 53 (TP53),extracellular signal-regulated kinase 1 (MAPK1),tumor necrosis factor (TNF),MAPK14,etc.【Conclusion】 PRRSV infection might cause autophagy,iron death and apoptosis,and the active components of the Yinhua Gancao decoction might resist PRRSV infection by regulating genes such as TP53,TNF,CASP3 and STAT3,and cell cycle and apoptosis signaling pathways.

Key words: Porcine reproductive and respiratory syndrome virus (PRRSV); GEO database; Yinhua Gancao decoction; UPLC-Q-TOF-MS

中图分类号:

S853.74

闫普普, 刘欣, 林冰, 刘佳丽, 夏瑾瑾, 黄永熙, 白若男, 刘国平, 杨小林, 郭利伟. 基于生物信息学探讨猪繁殖与呼吸综合征病毒感染特征及银花甘草汤防治机制[J]. 中国畜牧兽医, 2023, 50(10): 4196-4209.

YAN Pupu, LIU Xin, LIN Bing, LIU Jiali, XIA Jinjin, HUANG Yongxi, BAI Ruonan, LIU Guoping, YANG Xiaolin, GUO Liwei. Exploring of the Infection Characteristics of Porcine Reproductive and Respiratory Syndrome Virus and the Prevention and Control Mechanism of Yinhua Gancao Decoction Based on Bioinformatics[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 4196-4209.

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