基于GEO数据库与网络药理学探究公英散治疗奶牛乳房炎的作用机制  

Exploring the mechanism of Gongying powder in the treatment of dairy cow mastitis based on GEO database and network pharmacology

作　　者：吴权基 肖良贵 黄奔 WU Quanji;XIAO Lianggui;HUANG Ben(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Guangxi Zhuang Autonomous Region People’s Hospital,Guangxi Academy of Medical Sciences,Nanning 530021,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004 [2]广西医学科学院广西壮族自治区人民医院,南宁530021

出　　处：《黑龙江畜牧兽医》2025年第3期76-83,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(32160171)。

摘　　要：为了探究公英散治疗奶牛乳房炎的作用机制,试验在中药系统药理学数据库(TCMSP)、Symmap2数据库中筛选公英散中各组分的活性成分,利用Swiss Target Predicton在线工具预测其对应靶点;在GEO数据库中选择GSE15020数据集获取乳房炎相关差异表达基因;运用韦恩图在线作图平台得到公英散治疗奶牛乳房炎的交集靶点;对交集靶点进行蛋白互作网络分析及核心靶点预测;并对交集靶点进行GO富集分析和KEGG信号通路富集分析;构建公英散-活性成分-药物靶点-通路网络。设置空白(不加入细胞,只加入培养基和等体积溶剂)、对照(细胞不进行药物处理而只加入培养基和等体积溶剂)和诱导处理(细胞中分别加入培养基和150,200,250,300,500,1000μg/mL的公英散药液),每个处理6个重复,采用CCK8法筛选公英散药液质量浓度。根据筛选得到的公英散药液质量浓度设公英散低剂量组、中剂量组、高剂量组及空白组、模型组,采用Western-blot检测关键靶点蛋白表达水平。结果表明:共筛选出公英散药物活性成分75个,对应靶点729个,乳房炎差异表达基因281个,二者交集靶点42个;交集靶点蛋白互作网络分析度值最高的3个潜在靶点为IL-6、PTGS2、MMP9;相关通路主要涉及疟疾、糖尿病并发症的AGE-RAGE信号、脂质与动脉粥样硬化、TNF信号通路、PPAR信号通路和NF-κB通路等;公英散作用靶点度值排名前5位的是CA2、MMP9、EGFR、PTGS2、CA6。因此,选择IL-6、MMP9和PTGS23个关键靶点蛋白进行试验验证。与对照相比,150,200,250,300μg/mL的公英散药液对牛乳腺上皮细胞活力影响不显著(P>0.05),500,1000μg/mL的公英散药液对细胞活力具有极显著的抑制作用(P<0.01)。因此,选择150,200,250μg/mL的公英散药液(分别作为低、中、高剂量组)进行后续试验。与空白组相比,模型组IL-6、PTGS2和MMP9相对表达量极显著升高(P<0.01);与模型组相比,低剂量组IL-6In order to explore the mechanism of Gongying powder in the treatment of dairy cow mastitis,in this experiment,the active ingredients of each drug of Gongying powder were screened by using databases such as TCMSP and Symmap2 databases.The Swiss Target Predicton online tool was used to predict its corresponding targets.The GSE15020 dataset was selected in the GEO database to obtain the differentially expressed genes related to mastitis.The Venn diagram online mapping platform was used to obtain the intersection targets of Gongying powder in the treatment of dairy cow mastitis.Protein-protein interaction network analysis and core targets were predicted based on intersecting targets.GO enrichment analysis and KEGG signaling pathway enrichment analysis were performed on the intersecting targets.A network of Gongying powder-active ingredient-drug target-pathway was constructed.Blank group(no cells were added;only medium and equal volume solvent were added),control group(cells were not treated with drugs;only medium and equal volume solvent were added)and induction treatment groups(medium and 150,200,250,300,500,1000μg/mL of Gongying powder were added to the cells,respectively)were designed,with 6 replicates in each treatment;the CCK8 method was used to screen the mass concentration of Gongying powder.According to the liquid mass concentration of Gongying powder obtained from screening,the low-dose group,medium-dose group,high-dose group,cell blank group and model group were set up.Western-blot was used to detect the protein expression levels of key targets.The results showed that a total of 75 active pharmaceutical components of Gongying powder,729 corresponding targets,281 differential expressed genes of mastitis,and 42 intersecting targets were screened.The three potential targets with the highest analysis value of the intersecting target protein-protein interaction network were IL-6,PTGS2 and MMP9.The related pathways mainly involved malaria or diabetic complications-related AGE-RAGE signaling,lipid and atheroscle

关 键 词：GEO数据库 网络药理学 公英散 奶牛乳房炎 细胞试验验证 

分 类 号：S859[农业科学—临床兽医学]