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Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome

Intestinal ischemia and reperfusion injury is a model system of possible consequences of severe trauma and surgery, which might result into tissue dysfunction and organ failure. Neutrophils contribute to the injuries preceded by ischemia and reperfusion. However, the mechanisms by which intestinal i...

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Autores principales: Tahir, Muhammad, Arshid, Samina, Fontes, Belchor, Castro, Mariana S., Luz, Isabelle S., Botelho, Katyelle L. R., Sidoli, Simone, Schwämmle, Veit, Roepstorff, Peter, Fontes, Wagner
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6281993/
https://www.ncbi.nlm.nih.gov/pubmed/30555831
http://dx.doi.org/10.3389/fmolb.2018.00089
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author Tahir, Muhammad
Arshid, Samina
Fontes, Belchor
Castro, Mariana S.
Luz, Isabelle S.
Botelho, Katyelle L. R.
Sidoli, Simone
Schwämmle, Veit
Roepstorff, Peter
Fontes, Wagner
author_facet Tahir, Muhammad
Arshid, Samina
Fontes, Belchor
Castro, Mariana S.
Luz, Isabelle S.
Botelho, Katyelle L. R.
Sidoli, Simone
Schwämmle, Veit
Roepstorff, Peter
Fontes, Wagner
author_sort Tahir, Muhammad
collection PubMed
description Intestinal ischemia and reperfusion injury is a model system of possible consequences of severe trauma and surgery, which might result into tissue dysfunction and organ failure. Neutrophils contribute to the injuries preceded by ischemia and reperfusion. However, the mechanisms by which intestinal ischemia and reperfusion stimulate and activate circulating neutrophils is still not clear. In this work, we used proteomics approach to explore the underlying regulated mechanisms in Wistar rat neutrophils after ischemia and reperfusion. We isolated neutrophils from three different biological groups; control, sham laparotomy, and intestinal ischemia/reperfusion. In the workflow, we included iTRAQ-labeling quantification and peptide fractionation using HILIC prior to LC-MS/MS analysis. From proteomic analysis, we identified 2,045 proteins in total that were grouped into five different clusters based on their regulation trend between the experimental groups. A total of 417 proteins were found as significantly regulated in at least one of the analyzed conditions. Interestingly, the enzyme prediction analysis revealed that ischemia/reperfusion significantly reduced the relative abundance of most of the antioxidant and pro-survival molecules to cause more tissue damage and ROS production whereas some of the significantly up regulated enzymes were involved in cytoskeletal rearrangement, adhesion and migration. Clusters based KEGG pathways analysis revealed high motility, phagocytosis, directional migration, and activation of the cytoskeletal machinery in neutrophils after ischemia and reperfusion. Increased ROS production and decreased phagocytosis were experimentally validated by microscopy assays. Taken together, our findings provide a characterization of the rat neutrophil response to intestinal ischemia and reperfusion and the possible mechanisms involved in the tissue injury by neutrophils after intestinal ischemia and reperfusion.
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spelling pubmed-62819932018-12-14 Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome Tahir, Muhammad Arshid, Samina Fontes, Belchor Castro, Mariana S. Luz, Isabelle S. Botelho, Katyelle L. R. Sidoli, Simone Schwämmle, Veit Roepstorff, Peter Fontes, Wagner Front Mol Biosci Molecular Biosciences Intestinal ischemia and reperfusion injury is a model system of possible consequences of severe trauma and surgery, which might result into tissue dysfunction and organ failure. Neutrophils contribute to the injuries preceded by ischemia and reperfusion. However, the mechanisms by which intestinal ischemia and reperfusion stimulate and activate circulating neutrophils is still not clear. In this work, we used proteomics approach to explore the underlying regulated mechanisms in Wistar rat neutrophils after ischemia and reperfusion. We isolated neutrophils from three different biological groups; control, sham laparotomy, and intestinal ischemia/reperfusion. In the workflow, we included iTRAQ-labeling quantification and peptide fractionation using HILIC prior to LC-MS/MS analysis. From proteomic analysis, we identified 2,045 proteins in total that were grouped into five different clusters based on their regulation trend between the experimental groups. A total of 417 proteins were found as significantly regulated in at least one of the analyzed conditions. Interestingly, the enzyme prediction analysis revealed that ischemia/reperfusion significantly reduced the relative abundance of most of the antioxidant and pro-survival molecules to cause more tissue damage and ROS production whereas some of the significantly up regulated enzymes were involved in cytoskeletal rearrangement, adhesion and migration. Clusters based KEGG pathways analysis revealed high motility, phagocytosis, directional migration, and activation of the cytoskeletal machinery in neutrophils after ischemia and reperfusion. Increased ROS production and decreased phagocytosis were experimentally validated by microscopy assays. Taken together, our findings provide a characterization of the rat neutrophil response to intestinal ischemia and reperfusion and the possible mechanisms involved in the tissue injury by neutrophils after intestinal ischemia and reperfusion. Frontiers Media S.A. 2018-11-29 /pmc/articles/PMC6281993/ /pubmed/30555831 http://dx.doi.org/10.3389/fmolb.2018.00089 Text en Copyright © 2018 Tahir, Arshid, Fontes, Castro, Luz, Botelho, Sidoli, Schwämmle, Roepstorff and Fontes. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Tahir, Muhammad
Arshid, Samina
Fontes, Belchor
Castro, Mariana S.
Luz, Isabelle S.
Botelho, Katyelle L. R.
Sidoli, Simone
Schwämmle, Veit
Roepstorff, Peter
Fontes, Wagner
Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title_full Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title_fullStr Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title_full_unstemmed Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title_short Analysis of the Effect of Intestinal Ischemia and Reperfusion on the Rat Neutrophils Proteome
title_sort analysis of the effect of intestinal ischemia and reperfusion on the rat neutrophils proteome
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6281993/
https://www.ncbi.nlm.nih.gov/pubmed/30555831
http://dx.doi.org/10.3389/fmolb.2018.00089
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