High performance mass spectrometry based proteomics reveals enzyme and signaling pathway regulation in neutrophils during the early stage of surgical trauma.
In clinical conditions trauma is associated with high mortality and morbidity. Neutrophils play a key role in the development of multiple organ failure after trauma EXPERIMENTAL DESIGN: To have a detailed understanding of the neutrophil activation at primary stages after trauma, neutrophils are isolated from control and surgical trauma rats in this study. Extracted proteins are analyzed using nano liquid chromatography coupled with tandem mass spectrometry.



A total of 2924 rat neutrophil proteins are identified in our analysis, of which 393 are found differentially regulated between control and trauma groups. By using functional pathways analysis of the 190 proteins up-regulated in surgical trauma, we found proteins related to transcription initiation and protein biosynthesis. On the other hand, among the 203 proteins down-regulated in surgical trauma we found enrichment for proteins of the immune response, proteasome degradation and actin cytoskeleton. Overall, enzyme prediction analysis revealed that regulated enzymes are directly involved in neutrophil apoptosis, directional migration and chemotaxis. Our observations are then confirmed by in silico protein-protein interaction analysis.



Collectively, our results reveal that neutrophils drastically regulate their biochemical pathways after the early stages of surgical trauma, showing lower activity. This implies higher susceptibility of the trauma patients to infection and bystander tissues damage.
Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasília, Brazil.|Laboratory of Surgical Physiopathology (LIM-62), Faculty of Medicine, University of São Paulo, Brazil.
Samina
S
Arshid
Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasília, Brazil.|Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Muhammad
M
Tahir
Laboratory of Surgical Physiopathology (LIM-62), Faculty of Medicine, University of São Paulo, Brazil.
Belchor
B
Fontes
Laboratory of Surgical Physiopathology (LIM-62), Faculty of Medicine, University of São Paulo, Brazil.
Edna Frasson
EF
de Souza Montero
Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasília, Brazil.
Mariana S
MS
Castro
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Simone
S
Sidoli
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Peter
P
Roepstorff
Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasília, Brazil.
Wagner
W
Fontes
D000818:Animals; D017209:Apoptosis; D002851:Chromatography, High Pressure Liquid; D015536:Down-Regulation; D004798:Enzymes / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D007535:Isomerases / Q000032:analysis; D008297:Male; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism*; D010088:Oxidoreductases / Q000032:analysis; D060066:Protein Interaction Maps; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D015854:Up-Regulation; D014947:Wounds and Injuries / Q000378:metabolism / Q000473:pathology / Q000601:surgery
D004798:Enzymes; D020543:Proteome; D010088:Oxidoreductases; D007535:Isomerases
Inflammation; Neutrophil; Proteome; Surgery; Trauma