{"title":"大肠杆菌诱导牛乳腺的无标记定量蛋白质组学分析。","authors":"Li Zhou, Zhuoma Luoreng, Xingping Wang","doi":"10.2460/ajvr.24.12.0403","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To establish a model of Escherichia coli-induced bovine mammary gland inflammation and investigate alterations in protein expression in bovine mammary glands under different health conditions.</p><p><strong>Methods: </strong>6 Chinese Holstein cows were selected and randomly assigned to 2 groups. Bovine mammary glands were induced with either 105 CFU/mL E coli (n = 3) or 1 X PBS (n = 3), and label-free quantitative proteomics was applied to explore the effect of E coli on the expression of proteins in bovine mammary glands. Then, these data were combined with transcriptome sequencing data for integrative analysis.</p><p><strong>Results: </strong>A total of 944 differentially expressed proteins (DEPs) were identified; 588 proteins were upregulated and 356 were downregulated in mastitis tissues compared to healthy tissues. Gene Ontology functional annotations unveiled that the identified DEPs were predominantly enriched in processes, such as the single-organism metabolic process, extracellular region, and catalytic activity. According to Kyoto Encyclopedia of Genes and Genomes enrichment analysis, DEPs were prominently enriched in pathways related to type I diabetes mellitus, graft-versus-host disease, and peroxisome signaling pathways. InterPro analysis identified that the most significantly enriched functional structural domains were the epidermal growth factor-like calcium-binding domains, C-type lectin folds, and C-type lectins. By combining transcriptomic with proteomics data, 7 (cytochrome P450 family 51 subfamily A member 1, ethanolamine kinase 1, carbonic anhydrase 2, complement factor H, indoleamine 2,3-dioxygenase 1, CD5, and IL-33) hub genes (proteins) closely associated with mastitis development were identified.</p><p><strong>Conclusions: </strong>These findings offer a theoretical basis for understanding the molecular regulatory mechanisms underlying bovine mastitis.</p><p><strong>Clinical relevance: </strong>Our study presents a promising approach to understanding the mechanism of E coli infection, with a particular focus on the induction of bovine mastitis.</p>","PeriodicalId":7754,"journal":{"name":"American journal of veterinary research","volume":" ","pages":""},"PeriodicalIF":1.4000,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Label-free quantitative proteomics analysis of bovine mammary glands induced with Escherichia coli.\",\"authors\":\"Li Zhou, Zhuoma Luoreng, Xingping Wang\",\"doi\":\"10.2460/ajvr.24.12.0403\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To establish a model of Escherichia coli-induced bovine mammary gland inflammation and investigate alterations in protein expression in bovine mammary glands under different health conditions.</p><p><strong>Methods: </strong>6 Chinese Holstein cows were selected and randomly assigned to 2 groups. Bovine mammary glands were induced with either 105 CFU/mL E coli (n = 3) or 1 X PBS (n = 3), and label-free quantitative proteomics was applied to explore the effect of E coli on the expression of proteins in bovine mammary glands. Then, these data were combined with transcriptome sequencing data for integrative analysis.</p><p><strong>Results: </strong>A total of 944 differentially expressed proteins (DEPs) were identified; 588 proteins were upregulated and 356 were downregulated in mastitis tissues compared to healthy tissues. Gene Ontology functional annotations unveiled that the identified DEPs were predominantly enriched in processes, such as the single-organism metabolic process, extracellular region, and catalytic activity. According to Kyoto Encyclopedia of Genes and Genomes enrichment analysis, DEPs were prominently enriched in pathways related to type I diabetes mellitus, graft-versus-host disease, and peroxisome signaling pathways. 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引用次数: 0
摘要
目的：建立大肠杆菌诱导的牛乳腺炎症模型，探讨不同健康状态下牛乳腺蛋白表达的变化。方法：选取6头中国荷斯坦奶牛，随机分为2组。用105 CFU/mL大肠杆菌（n = 3）或1 X PBS （n = 3）诱导牛乳腺，采用无标记定量蛋白质组学方法研究大肠杆菌对牛乳腺蛋白表达的影响。然后，将这些数据与转录组测序数据结合进行整合分析。结果：共鉴定出944个差异表达蛋白（DEPs）；与健康组织相比，乳腺炎组织中有588种蛋白上调，356种蛋白下调。基因本体功能注释显示，所鉴定的DEPs主要富集于单生物代谢过程、细胞外区域和催化活性等过程。根据《京都基因和基因组百科全书》的富集分析，DEPs在与I型糖尿病、移植物抗宿主病和过氧化物酶体信号通路相关的通路中显著富集。InterPro分析发现，最显著富集的功能结构域是表皮生长因子样钙结合域、c型凝集素褶皱和c型凝集素。通过结合转录组学和蛋白质组学数据，鉴定出7个与乳腺炎发展密切相关的枢纽基因（蛋白）（细胞色素P450家族51亚家族A成员1、乙醇胺激酶1、碳酸酐酶2、补体因子H、吲哚胺2,3-双加氧酶1、CD5和IL-33）。结论：这些发现为理解奶牛乳腺炎的分子调控机制提供了理论基础。临床意义：我们的研究提出了一种有希望的方法来理解大肠杆菌感染的机制，特别关注牛乳腺炎的诱导。
Label-free quantitative proteomics analysis of bovine mammary glands induced with Escherichia coli.
Objective: To establish a model of Escherichia coli-induced bovine mammary gland inflammation and investigate alterations in protein expression in bovine mammary glands under different health conditions.
Methods: 6 Chinese Holstein cows were selected and randomly assigned to 2 groups. Bovine mammary glands were induced with either 105 CFU/mL E coli (n = 3) or 1 X PBS (n = 3), and label-free quantitative proteomics was applied to explore the effect of E coli on the expression of proteins in bovine mammary glands. Then, these data were combined with transcriptome sequencing data for integrative analysis.
Results: A total of 944 differentially expressed proteins (DEPs) were identified; 588 proteins were upregulated and 356 were downregulated in mastitis tissues compared to healthy tissues. Gene Ontology functional annotations unveiled that the identified DEPs were predominantly enriched in processes, such as the single-organism metabolic process, extracellular region, and catalytic activity. According to Kyoto Encyclopedia of Genes and Genomes enrichment analysis, DEPs were prominently enriched in pathways related to type I diabetes mellitus, graft-versus-host disease, and peroxisome signaling pathways. InterPro analysis identified that the most significantly enriched functional structural domains were the epidermal growth factor-like calcium-binding domains, C-type lectin folds, and C-type lectins. By combining transcriptomic with proteomics data, 7 (cytochrome P450 family 51 subfamily A member 1, ethanolamine kinase 1, carbonic anhydrase 2, complement factor H, indoleamine 2,3-dioxygenase 1, CD5, and IL-33) hub genes (proteins) closely associated with mastitis development were identified.
Conclusions: These findings offer a theoretical basis for understanding the molecular regulatory mechanisms underlying bovine mastitis.
Clinical relevance: Our study presents a promising approach to understanding the mechanism of E coli infection, with a particular focus on the induction of bovine mastitis.
期刊介绍：
The American Journal of Veterinary Research supports the collaborative exchange of information between researchers and clinicians by publishing novel research findings that bridge the gulf between basic research and clinical practice or that help to translate laboratory research and preclinical studies to the development of clinical trials and clinical practice. The journal welcomes submission of high-quality original studies and review articles in a wide range of scientific fields, including anatomy, anesthesiology, animal welfare, behavior, epidemiology, genetics, heredity, infectious disease, molecular biology, oncology, pharmacology, pathogenic mechanisms, physiology, surgery, theriogenology, toxicology, and vaccinology. Species of interest include production animals, companion animals, equids, exotic animals, birds, reptiles, and wild and marine animals. Reports of laboratory animal studies and studies involving the use of animals as experimental models of human diseases are considered only when the study results are of demonstrable benefit to the species used in the research or to another species of veterinary interest. Other fields of interest or animals species are not necessarily excluded from consideration, but such reports must focus on novel research findings. Submitted papers must make an original and substantial contribution to the veterinary medicine knowledge base; preliminary studies are not appropriate.
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