基于网络药理学探讨黄芩提取物防治猪链球菌2型感染的作用机制

doi:10.16431/j.cnki.1671-7236.2023.12.038

摘要/Abstract

摘要： 【目的】探索黄芩提取物防治猪链球菌2型(Streptococcus suis type 2,SS2)感染的潜在作用机制。【方法】利用GEO数据库收集SS2感染不同细胞的转录组数据,通过GEO2R在线软件筛选差异表达基因,并利用STRING数据库与Cytoscape v3.8.2软件构建蛋白互作(PPI)网络并进行拓扑分析,通过GO功能和KEGG通路富集分析筛选SS2感染机体后核心蛋白、生物功能和信号转导途径变化。采用液相色谱质谱联用(LC-MS)分析并鉴定黄芩提取物的组成成分,利用Swiss ADME和SwissTargetPredication数据库筛选黄芩提取物的活性成分并预测其作用靶点;运用网络药理学方法将疾病靶点与黄芩提取物有效成分作用靶点相结合,进一步明确黄芩提取物抗SS2感染机制。通过分子对接和实时荧光定量PCR方法验证网络药理学预测结果的可行性。【结果】GEO数据库的2组基因芯片共鉴定出1 308个SS2感染差异表达基因,PPI分析筛选出核心基因70个,GO功能富集分析显示显著富集条目主要涉及细胞因子介导的信号通路和对细胞因子刺激的反应等,KEGG通路富集分析涉及TNF、PI3K-Akt、JAK-STAT等信号转导途径。黄芩提取物共包括31个有效成分,包括汉黄芩素和黄芩苷等,其可能影响VEGFA、TNF和PPARG等核心靶点,涉及炎症反应、细胞凋亡和促进血管新生等多个生物功能和信号通路。分子对接结果显示,黄芩提取物中汉黄芩素和黄芩苷与TNF、PPARG和VEGFA有较好的结合能。实时荧光定量PCR结果显示,SS2感染后猪肺泡巨噬细胞(PAMs)细胞中CA9、PPARG、JUN、SCD、JAK3和VEGFA基因mRNA水平均显著上调(P＜0.05),黄芩提取物干预后,上述基因mRNA表达水平均显著下降(P＜0.05)。【结论】SS2感染机体后涉及细胞因子介导的生物过程、TNF等信号通路发生变化,黄芩提取物有效成分可能通过AMPK、PI3K-Akt、GFR、TNF等信号通路共同参与机体凋亡调控、炎症反应、血管生成等生物过程从而抗SS2感染。

关键词: 猪链球菌2型; 黄芩提取物; 液质联用; 网络药理学; 分子对接

Abstract: 【Objective】 The aim of this study was to explore the potential mechanism of Scutellaria baicalensis Georgi extract in preventing and treating Streptococcus suis type 2 (SS2) infection.【Method】 The transcriptome data of SS2-infected cells were collected by GEO database,and differentially expressed genes were screened by GEO2R online software.The protein-protein interaction (PPI) inetwork was constructed using STRING database and Cytoscape v3.8.2 software,and topological analysis was performed.The changes of core protein,biological function and signal transduction pathway after SS2 infection were screened by GO function and KEGG pathway enrichment analysis.Liquid chromatography mass spectrometry (LC-MS) was used to analyze and identify the components of Scutellaria baicalensis Georgi extract,and Swiss ADME and SwissTargetPredication databases were used to screen the active components of Scutellaria baicalensis Georgi extract and predict its target.The mechanism of anti-SS2 infection of Scutellaria baicalensis Georgi extract was further defined by combining the disease target with the active ingredient target by network pharmacology.Molecular docking and Real-time quantitative PCR were used to verify the feasibility of the network pharmacological prediction results.【Result】 1 308 differential genes of SS2 infection were identified by two sets of gene chips provided by GEO database,and 70 core genes were screened by PPI analysis.GO functional enrichment analysis showed that significant enrichment items mainly involved cytokine-mediated signaling pathways and responses to cytokine stimulation.KEGG pathway enrichment analysis involved the TNF,PI3K-Akt,JAK-STAT and other signal transduction pathways.A total of 31 active components of Scutellaria baicalensis Georgi extract,including baicalin and baicalin,etc.,which might affect core targets such as VEGFA,TNF and PPARG,involving multiple biological functions and signaling pathways such as inflammatory response,apoptosis and promotion of angiogenesis.Molecular docking results showed that baicalin and baicalin of Scutellaria baicalensis Georgi extract had good binding energy with TNF,PPARG and VEGFA.Real-time quantitative PCR results showed that the mRNA levels of CA9,PPARG,JUN,SCD,JAK3 and VEGFA genes in PAMs cells were significantly up-regulated after SS2 infection (P＜0.05),and the mRNA expression levels of these genes were significantly decreased after the intervention of Scutellaria baicalensis Georgi extract (P＜0.05).【Conclusion】 After SS2 infection,cytokine mediated biological processes,TNF and other signaling pathways changed and the active components of Scutellaria baicalensis Georgi extract might jointly participate in apoptosis regulation,inflammatory response,angiogenesis and other biological processes through AMPK,PI3K-Akt,GFR and TNF signaling pathways,thereby preventing SS2 infection.

Key words: Streptococcus suis type 2; Scutellaria baicalensis Georgi extract; LC-MS; network pharmacology; molecular docking

中图分类号:

S855.1^＋1

刘佳丽, 闫普普, 刘恋, 朱君, 夏瑾瑾, 黄永熙, 白若男, 郭利伟, 刘国平, 杨小林. 基于网络药理学探讨黄芩提取物防治猪链球菌2型感染的作用机制[J]. 中国畜牧兽医, 2023, 50(12): 5170-5185.

LIU Jiali, YAN Pupu, LIU Lian, ZHU Jun, XIA Jinjin, HUANG Yongxi, BAI Ruonan, GUO Liwei, LIU Guoping, YANG Xiaolin. Exploring the Mechanism of Scutellaria baicalensis Georgi Extract in Preventing and Treating Streptococcus suis Type 2 Infection Based on Network Pharmacology[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5170-5185.

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