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(Journal of Leukocyte Biology. 2001;69:241-247.)
© 2001 by Society for Leukocyte Biology

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

Keigo Nakatani, Seiichiro Takeshita, Hiroshi Tsujimoto, Youichi Kawamura and Isao Sekine

Department of Pediatrics, National Defense Medical College, Tokorozawa, Saitama, Japan

Correspondence: Seiichiro Takeshita, M.D., Ph.D., Department of Pediatrics, National Defense Medical College, Namiki 3-2, Tokorozawa, Saitama 359-8513, Japan. E-mail: peditake{at}ndmc.ac.jp

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 {alpha}-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.

Key Words: elastase • ulinastatin • ONO-5046




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