Anoxia-reoxygenation-induced, neutrophil-mediated endothelial cell injury: role of elastase.
The aim of this study was to assess the role of neutrophilic elastase in anoxia-reoxygenation-induced, neutrophil-mediated injury to microvascular endothelium. Cultured bovine microvascular endothelial cells were grown to confluence and labeled with 51Cr. The endothelial cells were exposed to a 30-min period of anoxia and subsequently reoxygenated. Endothelial cell injury, quantitated as 51Cr release and cell detachment, was determined 8 h after reoxygenation. Addition of neutrophils upon reoxygenation enhanced the anoxia-reoxygenation-induced increase in 51Cr release and cell detachment. The neutrophil-mediated injury was associated with elastase release from the neutrophils. Four agents were used to inhibit neutrophilic elastase activity: Eglin C, methoxysuccunyl-Ala2-Pro-Val-CH2Cl, L658,758, and a monoclonal antibody against neutrophilic elastase. All elastase inhibitors attenuated the neutrophil-mediated endothelial cell detachment but not 51Cr release. Addition of purified human neutrophilic elastase, at a level that mimicked the release from neutrophils, increased cell detachment in endothelial cells exposed to anoxia-reoxygenation but did not affect 51Cr release. Our results indicate that elastase plays an important role in anoxia-reoxygenation-induced, neutrophil-mediated endothelial cell dysfunction.
Inauen
W
W
Department of Physiology and Biophysics, Louisiana State University Medical Center, Shreveport 71130.
Granger
D N
DN
Meininger
C J
CJ
Schelling
M E
ME
Granger
H J
HJ
Kvietys
P R
PR
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D004730:Endothelium, Vascular / Q000473:pathology*; D000860:Hypoxia / Q000473:pathology* / Q000503:physiopathology; D009504:Neutrophils / Q000201:enzymology / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology*; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors / Q000494:pharmacology / Q000502:physiology*
D010196:Pancreatic Elastase; D010100:Oxygen