{"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. 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":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"American journal of veterinary research","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.2460/ajvr.24.12.0403","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/6/1 0:00:00","PubModel":"Print","JCR":"Q2","JCRName":"VETERINARY SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
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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