Introduction Circulatory shock is a frequent cause of death and one of the most important unresolved medical problems. Reduction of blood supply to the intestine during ischemia disrupts the mucosal epithelial barrier allowing inflammatory materials in the lumen of the intestine, including digestive enzymes, to cross the intestinal wall. The diffusion of digestive enzymes in the circulation generates a strong inflammatory reaction. Recent studies have demonstrated that the uncontrolled proteolytic activity causes cleavage of important membrane receptors on the endothelial cells. Thus, understanding the contribution of degrading proteases to circulatory shock may be essential to elucidate the mechanisms and the pathways leading to multi-organ failure and to detect potential biomarkers and/or therapeutic targets. Methods Peptides and low molecular weight proteins were isolated from plasma using a two-step enrichment method (ultra filtration followed by solvent precipitation) and analyzed by a label-free quantitative peptidomic approach based on nano-LC-LTQ-Orbitrap Velos MS/MS Results and Conclusions This study was aimed at comparing the peptidome of plasma samples from healthy and hemorrhagic-shock rats to verify the possible role of uncontrolled proteolytic activity. Peptides were considered as differentially expressed if they were present only in healthy (C) or hemorrhagic (HS) groups or showed significant fold change differences (>1.5 or <0.67). 256 peptides were up-regulated or present only in HS group, while 60 peptides were down-regulated in HS group or were present only in C group. Our results strongly suggest that there is an increase in protease activity in the systemic circulation following hemorrhagic shock. Further analysis, conducted to determine which proteases potentially derived from the intestine may contribute to the uncontrolled degradative activity, reveals a significant increase of chymotryptic activity in post shock plasma compared to controls.
Proteomic analysis of plasma peptidome in healthy and hemorrhagic-shock rats / E. Maffioli, F. Santagata, F. Aletti, S. Nonnis, G. Schmid Schönbein, A. Negri, G. Tedeschi. ((Intervento presentato al convegno 58 National Meeting of the Italian Society of Biochemistry and Molecular Biology tenutosi a Urbino nel 2015.
Proteomic analysis of plasma peptidome in healthy and hemorrhagic-shock rats
F. Santagata;S. Nonnis;A. Negri;G. Tedeschi
2015
Abstract
Introduction Circulatory shock is a frequent cause of death and one of the most important unresolved medical problems. Reduction of blood supply to the intestine during ischemia disrupts the mucosal epithelial barrier allowing inflammatory materials in the lumen of the intestine, including digestive enzymes, to cross the intestinal wall. The diffusion of digestive enzymes in the circulation generates a strong inflammatory reaction. Recent studies have demonstrated that the uncontrolled proteolytic activity causes cleavage of important membrane receptors on the endothelial cells. Thus, understanding the contribution of degrading proteases to circulatory shock may be essential to elucidate the mechanisms and the pathways leading to multi-organ failure and to detect potential biomarkers and/or therapeutic targets. Methods Peptides and low molecular weight proteins were isolated from plasma using a two-step enrichment method (ultra filtration followed by solvent precipitation) and analyzed by a label-free quantitative peptidomic approach based on nano-LC-LTQ-Orbitrap Velos MS/MS Results and Conclusions This study was aimed at comparing the peptidome of plasma samples from healthy and hemorrhagic-shock rats to verify the possible role of uncontrolled proteolytic activity. Peptides were considered as differentially expressed if they were present only in healthy (C) or hemorrhagic (HS) groups or showed significant fold change differences (>1.5 or <0.67). 256 peptides were up-regulated or present only in HS group, while 60 peptides were down-regulated in HS group or were present only in C group. Our results strongly suggest that there is an increase in protease activity in the systemic circulation following hemorrhagic shock. Further analysis, conducted to determine which proteases potentially derived from the intestine may contribute to the uncontrolled degradative activity, reveals a significant increase of chymotryptic activity in post shock plasma compared to controls.
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