Verocytotoxin-induced apoptosis of human microvascular endothelial cells

Citation
Ahjm. Pijpers et al., Verocytotoxin-induced apoptosis of human microvascular endothelial cells, J AM S NEPH, 12(4), 2001, pp. 767-778
Citations number
46
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Urology & Nephrology","da verificare
Journal title
JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY
ISSN journal
1046-6673 → ACNP
Volume
12
Issue
4
Year of publication
2001
Pages
767 - 778
Database
ISI
SICI code
1046-6673(200104)12:4<767:VAOHME>2.0.ZU;2-W
Abstract
The pathogenesis of the epidemic form of hemolytic uremic syndrome is chara cterized by endothelial cell damage. In this study, the role of apoptosis i n verocytotoxin (VT)mediated endothelial cell death in human glomerular mic rovascular endothelial cells (GMVEC), human umbilical vein endothelial cell s, and foreskin microvascular endothelial cells (FMVEC) was investigated. V T induced apoptosis in GMVEC and human umbilical vein endothelial cells whe n the cells were prestimulated with the inflammatory mediator tumor necrosi s factor-alpha (TNF-alpha). FMVEC displayed strong binding of VT and high s usceptibility to VT under basal conditions, which made them suitable for th e study of VT-induced apoptosis without TNF-alpha interference. On the basi s of functional (flow cytometry and immunofluorescence microscopy using FIT C-conjugated annexin V and propidium iodide), morphologic (transmission ele ctron microscopy), and molecular (agarose gel electrophoresis of cellular D NA fragments) criteria, it was documented that VT induced programmed cell d eath in microvascular endothelial cells in a dose- and time-dependent manne r. Furthermore, whereas partial inhibition of protein synthesis by VT was a ssociated with a considerable number of apoptotic cells, comparable inhibit ion of protein synthesis by cycloheximide was not. This suggests that addit ional pathways, independent of protein synthesis inhibition, may be involve d in VT-mediated apoptosis in microvascular endothelial cells. Specific inh ibition of caspases by Ac-Asp-Glu-Val-Asp-CHO, but not by Ac-Tyr-Val-Ala-As p-CHO, was accompanied by inhibition of VT-induced apoptosis in FMVEC and T NF-alpha -treated GMVEC. These data indicate that VT can induce apoptosis i n human microvascular endothelial cells.