Leucocyte elastase-mediated release of von Willebrand factor from cultured endothelial cells.
The aim of this study was to investigate a possible activation of human endothelial cells in monolayer culture by a purified neutrophil-derived proteinase, i.e. elastase. Cells were isolated from human umbilical cord veins, and incubated either in primary culture or after two passages, in the presence of various concentrations of this proteinase. Although a lack of prostacyclin formation was noted, elastase induced a large release of von Willebrand factor (vWf) in a concentration-dependent manner. Thus, upon incubation with 1 microgram.ml-1 elastase for 30 min, 70 mU.ml-1 vWf were detected in the incubation medium, as compared to 5 mU.ml-1 for control. Using cells in primary culture, a fivefold higher concentration of vWf was recovered following incubation with 10 micrograms.ml-1 elastase than with 0.5 IU.ml-1 thrombin. This effect was linked to the enzymatic activity of elastase and not to its cationic charge, as deduced from the inhibition by eglin C, and the lack of effect of the phenylmethylsulphonyl fluoride (PMSF) treated proteinase. We conclude that vWf release was not due to cell activation, since cytoplasmic calcium mobilization was absent, and inhibition of protein kinase C did not modify the response. In fact, this release was the consequence of cell damage, since concentrations of vWf recovered correlated with cell lysis. These results support the hypothesis that the high level of plasma vWf in patients with sepsis or adult respiratory distress syndrome could result from damage to the endothelial cells by elastase released from activated neutrophils.
Chignard
M
M
Unité de Pharmacologie Cellulaire, Unité associée IP-INSERM 285, Institut Pasteur, Paris, France.
Balloy
V
V
Renesto
P
P
D002118:Calcium / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D019272:Leukocyte Elastase; D010196:Pancreatic Elastase / Q000008:administration & dosage / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011453:Prostaglandins / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology; D014841:von Willebrand Factor / Q000187:drug effects* / Q000378:metabolism
D011453:Prostaglandins; D014841:von Willebrand Factor; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase; D002118:Calcium