title: Changes in protein expression profiles in bovine endometrial epithelial cells exposed to E. coli LPS challenge.
abstract: E. coli is one of the most frequently involved bacteria in uterine diseases. Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria involved in pathogenic processes leading to post-partum metritis and endometritis in cattle. It also causes inflammation of the endometrium. The increase of cell proliferation by LPS is part of the inflammatory process. The aim of this study was to investigate possible changes in protein expression in relation to the proliferative response of bEECs after challenge with E. coli-LPS. In vitro culture of bEECs was performed from cow genital tracts collected at a slaughterhouse. In passage 5, bEECs from each of 9 cows (3 series of 3 cows) were exposed to 0, 8, and 16 μg ml-1LPS for 72 h. At time 0 and 72 h later, attached cells/living cells were counted and for each time and LPS dosage, cells were frozen for proteomic analyses. All samples from the 3 series were analyzed by 2-D gel electrophoresis coupled to MALDI-TOF/TOF mass spectrometry. The samples from the first series were subjected to shotgun nLC-MS/MS analysis. From the whole differential proteomics analysis, 38 proteins were differentially expressed (p < 0.05 to p < 0.001) following exposure to LPS. Among them, twenty-eight were found to be up-regulated in the LPS groups in comparison to control groups and ten were down-regulated. Differentially expressed proteins were associated with cell proliferation and apoptosis, transcription, destabilization of cell structure, oxidative stress, regulation of histones, allergy and general cell metabolism pathways. The de-regulations induced by LPS were consistent with the proliferative phenotype and indicated strong alterations of several cell functions. In addition, some of the differentially expressed proteins relates to pathways activated at the time of implantation. The specific changes induced through those signals may have negative consequences for the establishment of pregnancy.
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli studi di Milano, Milano, Italy.
authors.forename: Cristian
authors.initials: C
authors.lastname: Piras
authors.affiliation: Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, SLU, Sweden.
authors.forename: Yongzhi
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli studi di Milano, Milano, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Soggiu
authors.affiliation: Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, SLU, Sweden and Rajamangala University of Srivijaya (RMUTSV), Thungyai, Thailand.
authors.forename: Metasu
authors.initials: M
authors.lastname: Chanrot
authors.affiliation: Proteomics and Metabonomics Unit Fondazione Santa Lucia - IRCCS, Rome, Italy.
authors.forename: Viviana
authors.initials: V
authors.lastname: Greco
authors.affiliation: Istituto di Biochimica e Biochimica Clinica, Università Cattolica, Roma, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Urbani
authors.affiliation: INRA, Biologie du Développement et Reproduction, 78350, Jouy en Josas, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Charpigny
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli studi di Milano, Milano, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Bonizzi
authors.affiliation: Dipartimento di Medicina Veterinaria, Università degli studi di Milano, Milano, Italy and Istituto Sperimentale Italiano L. Spallanzani, Milano and TechnologieS srl, via Celoria 10, 20133 Milano, Italy. paola.roncada@istitutospallanzani.it paola.roncada@gmail.com.
authors.forename: Paola
authors.identifier: http://orcid.org/0000-0002-0114-5872
authors.initials: P
authors.lastname: Roncada
authors.affiliation: Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, SLU, Sweden.
authors.forename: Patrice
authors.initials: P
authors.lastname: Humblot
mesh_terms: D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000235:genetics; D004717:Endometrium / Q000378:metabolism*; D004734:Energy Metabolism; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism*; D004926:Escherichia coli / Q000276:immunology; D005260:Female; D008070:Lipopolysaccharides / Q000009:adverse effects* / Q000276:immunology; D053858:Metabolic Networks and Pathways; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D020543:Proteome; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D057188:Workflow
chemical_list: D008070:Lipopolysaccharides; D020543:Proteome