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Titolo:
Influence of intercellular junctions on endothelin secretion of human umbilical vein endothelial cells in vitro
Autore:
Kiessling, F; Becker, D; Ullisch, EV; Kubler, W; Haller, C;
Indirizzi:
Univ Heidelberg, Med Klin, Abt 3, D-69115 Heidelberg, Germany Univ Heidelberg Heidelberg Germany D-69115 , D-69115 Heidelberg, Germany
Titolo Testata:
BASIC RESEARCH IN CARDIOLOGY
fascicolo: 4, volume: 95, anno: 2000,
pagine: 299 - 307
SICI:
0300-8428(200008)95:4<299:IOIJOE>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-KINASE-C; SHEAR-STRESS; IN-VITRO; CADHERINS; CA2+; GENE; ADHESION; CALCIUM;
Keywords:
endothelial junctions; calcium; endothelin; cadherins; von Willebrand factor;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
21
Recensione:
Indirizzi per estratti:
Indirizzo: Haller, C Univ Heidelberg, Med Klin, Abt 3, Bergheimer Str 58, D-69115 Heidelberg, Germany Univ Heidelberg Bergheimer Str 58 Heidelberg Germany D-69115 ny
Citazione:
F. Kiessling et al., "Influence of intercellular junctions on endothelin secretion of human umbilical vein endothelial cells in vitro", BAS R CARD, 95(4), 2000, pp. 299-307

Abstract

The endothelium plays a pivotal role in the rheological regulation of blood flow by the secretion of vasoactive factors. The interaction between shear forces and the endothelium is determined by the mechanical properties of the endothelial cell layer which are associated with intercellular junctions. Cell-cell contacts could therefore modulate the secretion of vasocative factors in response to rheological stimuli. We investigated the relationship between intercellular junctions and the secretion of the vasoconstrictor peptide endothelin and the coagulation co-factor von Willebrand factor (vWF). Human umbilical vein endothelial cells (HUVECs) were used as in vitro endothelial model system. Intercellular junctions were reversibly disrupted by calcium chelation or hypertonic stress; alternatively, the formation of intercellular junctions was inhibited by culturing the cells in suspension or by plating them in the presence of an inhibitory anti-VE-cadherin antibody. The opening of intercellular junctions was verified by assessing transmonolayer electrical resistance (TMR) and immunofluorescence morphology. The concentration of endothelin and vWF was measured in the cell culture supernatants using specific ELISAs. The secretion of endothelin was inhibited by EGTA (5 mM) and stimulated by incubation with tumor necrosis factor alpha (TNF alpha, 40 ng/ml). Treatment with hypertonic medium(glycerol, 1200 mosmol/l) for 10 minutes opened intercellular junctions and markedly reduced thesecretion of endothelin. HUVECs in suspension culture did not secrete endothelin and failed to respond to TNF alpha, but readily resumed these functions upon forming a new monolayer on plastic. The reconstitution of intercellular junctions after suspension culture could be inhibited using a specific anti-VE-cadherin antibody. This antibody, but not a non-specific anti-human-Iga antibody reduced endothelin secretion. The secretion of von Willebrand Factor was less dependent on intercellular junctions. The opening of intercellular junctions did not induce cell death, since the cells continued to exclude trypan blue. The results of this study suggest a novel and potentially pathophysiologically/clinically relevant correlation between intercellular junctions and the secretion of endothelin in endothelial cells.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 20/09/20 alle ore 00:07:34