Cationic neutrophil proteins increase transendothelial albumin movement.
Neutrophils play a role in the development of pulmonary edema in many models of the adult respiratory distress syndrome, but the mechanism of their action is not completely understood. We asked whether two neutrophil secretory products, human neutrophil cationic protein (NCP) and human neutrophil elastase (HNE), would nonenzymatically alter the movement of albumin across a cultured endothelial monolayer. Both enzymes were inactivated by heating before use. HNE was additionally enzymatically inactivated with a chloromethylketone oligopeptide (CMK) inhibitor and with alpha 1-proteinase inhibitor (alpha 1-PI). Heated NCP, heated HNE, and CMK-complexed HNE all increased transendothelial albumin transfer. The cation protamine also increased albumin transfer across the endothelium and this increase was blocked by heparin. Alpha 1-PI and fetal bovine serum also prevented the cationic proteins from increasing albumin transfer. Using the release of lactate dehydrogenase as a marker of cytotoxicity, heated HNE was toxic to endothelial cells, heated NCP had only minimal toxicity, and protamine had no toxicity. Changes in endothelial cell shape with gap formation was seen after exposure to both heated HNE and heated NCP. Both the cytotoxicity associated with heated HNE and the cell shape changes associated with heated NCP and heated HNE could be blocked by heparin. These results suggest that in addition to neutrophil proteases and reactive O2 molecules, neutrophil-derived cationic proteins can directly and nonenzymatically contribute to edema formation during acute inflammation.
D023181:Antimicrobial Cationic Peptides; D001696:Biomechanical Phenomena; D001790:Blood Physiological Phenomena; D001798:Blood Proteins / Q000502:physiology*; D002199:Capillary Permeability; D004563:Electrochemistry; D004727:Endothelium / Q000378:metabolism; D006801:Humans; D009504:Neutrophils / Q000201:enzymology; D010196:Pancreatic Elastase / Q000097:blood; D011479:Protamines / Q000494:pharmacology; D011651:Pulmonary Artery / Q000378:metabolism*; D012709:Serum Albumin / Q000378:metabolism*
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D011479:Protamines; D012709:Serum Albumin; C030509:neutrophil basic proteins; D010196:Pancreatic Elastase