Inhibitory effect of serine protease inhibitors on neutrophil-mediated endothelial cell injury

• Published in: Journal of leukocyte biology Vol. 69; no. 2; pp. 241 - 247
• Main Authors: Nakatani, Keigo, Takeshita, Seiichiro, Tsujimoto, Hiroshi, Kawamura, Youichi, Sekine, Isao
• Format: Journal Article
• Language: English
• Published: United States Society for Leukocyte Biology 01.02.2001
• Subjects: aprotinin; argatroban; Cells, Cultured; Drug Combinations; elastase; Endothelium, Vascular - drug effects; Endothelium, Vascular - enzymology; Endothelium, Vascular - immunology; Endothelium, Vascular - pathology; Flow Cytometry; gabexate mesilate; Glycine - analogs & derivatives; Glycine - pharmacology; Glycoproteins - pharmacology; Humans; Interferon-gamma - pharmacology; Interleukin-1 - pharmacology; Intracellular Fluid - drug effects; Intracellular Fluid - enzymology; Leukocyte Elastase - antagonists & inhibitors; Leukocyte Elastase - metabolism; Leukocyte Elastase - physiology; Lipopolysaccharides - pharmacology; Microscopy, Confocal; N-Formylmethionine Leucyl-Phenylalanine - pharmacology; nafamostat mesilate; Neutrophil Activation - immunology; Neutrophils - drug effects; Neutrophils - enzymology; Neutrophils - immunology; ONO‐5046; serine proteinase inhibitors; Serine Proteinase Inhibitors - pharmacology; Sulfonamides - pharmacology; Tetradecanoylphorbol Acetate - pharmacology; Trypsin Inhibitors - pharmacology; Tumor Necrosis Factor-alpha - pharmacology; ulinastatin; Umbilical Veins
• ISSN: 0741-5400; 1938-3673
• DOI: 10.1189/jlb.69.2.241

To investigate the inhibitory effect of serine protease inhibitors (SPI) on neutrophil‐mediated endothelial cell (EC) injury, we analyzed the in vitro cytotoxicity of radiolabeled human umbilical vein EC (HUVEC) mediated by neutrophils in the presence of SPI. The EC injury was inhibited dose‐dependently by urinary trypsin inhibitor (ulinastatin, UTI) and ONO‐5046, which have the ability to inactivate neutrophil elastase, but not by gabexate mesilate, nafamostat mesilate, aprotinin, and argatroban, which have no ability to inactivate neutrophil elastase. In addition, when UTI and ONO‐5046 were added to the tumor necrosis factor α‐primed neutrophils alone, they showed a dose‐dependent inhibition of the intracellular elastase activity, but the other SPI did not, for either flow cytometry or confocal microscopy. Therefore, UTI and ONO‐5046 may protect EC against the neutrophil‐mediated injury not only by inactivating the extracellular elastase secreted by neutrophils, but also by acting directly on neutrophils and suppressing the production and secretion of activated elastase from them.