title: Protease activity increases in plasma, peritoneal fluid, and vital organs after hemorrhagic shock in rats.
abstract: Hemorrhagic shock (HS) is associated with high mortality. A severe decrease in blood pressure causes the intestine, a major site of digestive enzymes, to become permeable - possibly releasing those enzymes into the circulation and peritoneal space, where they may in turn activate other enzymes, e.g. matrix metalloproteinases (MMPs). If uncontrolled, these enzymes may result in pathophysiologic cleavage of receptors or plasma proteins. Our first objective was to determine, in compartments outside of the intestine (plasma, peritoneal fluid, brain, heart, liver, and lung) protease activities and select protease concentrations after hemorrhagic shock (2 hours ischemia, 2 hours reperfusion). Our second objective was to determine whether inhibition of proteases in the intestinal lumen with a serine protease inhibitor (ANGD), a process that improves survival after shock in rats, reduces the protease activities distant from the intestine. To determine the protease activity, plasma and peritoneal fluid were incubated with small peptide substrates for trypsin-, chymotrypsin-, and elastase-like activities or with casein, a substrate cleaved by multiple proteases. Gelatinase activities were determined by gelatin gel zymography and a specific MMP-9 substrate. Immunoblotting was used to confirm elevated pancreatic trypsin in plasma, peritoneal fluid, and lung and MMP-9 concentrations in all samples after hemorrhagic shock. Caseinolytic, trypsin-, chymotrypsin-, elastase-like, and MMP-9 activities were all significantly (p<0.05) upregulated after hemorrhagic shock regardless of enteral pretreatment with ANGD. Pancreatic trypsin was detected by immunoblot in the plasma, peritoneal space, and lungs after hemorrhagic shock. MMP-9 concentrations and activities were significantly upregulated after hemorrhagic shock in plasma, peritoneal fluid, heart, liver, and lung. These results indicate that protease activities, including that of trypsin, increase in sites distant from the intestine after hemorrhagic shock. Proteases, including pancreatic proteases, may be shock mediators and potential targets for therapy in shock.
authors.affiliation: Department of Bioengineering, The Institute of Engineering in Medicine, University of California, San Diego, La Jolla, California, United States of America. aaltshul@ucsd.edu
authors.forename: Angelina E
authors.initials: AE
authors.lastname: Altshuler
authors.forename: Alexander H
authors.initials: AH
authors.lastname: Penn
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Yang
authors.forename: Ga-Ram
authors.initials: GR
authors.lastname: Kim
authors.forename: Geert W
authors.initials: GW
authors.lastname: Schmid-Schönbein
mesh_terms: D000818:Animals; D001202:Ascitic Fluid / Q000201:enzymology*; D001550:Benzamidines; D001826:Body Fluids / Q000201:enzymology*; D001921:Brain / Q000187:drug effects / Q000201:enzymology; D006146:Guanidines / Q000494:pharmacology; D015151:Immunoblotting; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008168:Lung / Q000187:drug effects / Q000201:enzymology; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D009206:Myocardium / Q000201:enzymology; D010529:Peritoneal Cavity; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D057057:Serine Proteases / Q000097:blood* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D012771:Shock, Hemorrhagic / Q000201:enzymology* / Q000503:physiopathology* / Q000517:prevention & control; D013379:Substrate Specificity; D014357:Trypsin / Q000378:metabolism
chemical_list: D001550:Benzamidines; D006146:Guanidines; D061965:Matrix Metalloproteinase Inhibitors; D015842:Serine Proteinase Inhibitors; D009195:Peroxidase; D057057:Serine Proteases; D014357:Trypsin; D020780:Matrix Metalloproteinase 9; C032855:nafamostat