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Effect of unfractionated heparin, enoxaparin and sulodexide on the relations between secretion and expression of OPG, RANKL and vWF in HUVEC
open access
Abstract
Heparin modulates function of vascular endothelium. We studied the effects of unfractionated heparin (UFH) vs. enoxaparin vs. sulodexide on the levels and gene expression of osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kB Ligand (RANKL) and von Willebrand factor (vWF) in Human Umbilical Vein Endothelial Cells (HUVEC) culture. HUVEC were isolated from human umbilical vein by a standard method. The supernatant concentrations (ELISA) and gene expression (Real Time-PCR) of OPG, RANKL and vWF in HUVEC were determined after incubation with various concentrations of UFH, enoxaparin and sulodexide for up to 16 hours. In control HUVEC strong positive correlation between OPG and vWF levels was observed, whereas sRANKL negatively correlated with OPG and vWF levels. Only in control HUVEC a negative correlation between the supernatant level of vWF and its gene expression was found. Already the lowest concentration of UFH caused 2.5-fold increase in OPG gene expression while higher UFH concentrations substantially increased RANKL mRNA level. A negative correlation between the OPG and sRANKL concentration was noticed in supernatant HUVEC which were incubated with enoxaparine. In conclusion, the observed interrelationships between OPG, RANKL and vWF levels in unstimulated HUVEC support the presumption of the pathophysiological links between these proteins. Of the tested heparin formulas UFH seems to be the most potent in altering the OPG, RANKL and vWF axis.
Abstract
Heparin modulates function of vascular endothelium. We studied the effects of unfractionated heparin (UFH) vs. enoxaparin vs. sulodexide on the levels and gene expression of osteoprotegerin (OPG), Receptor Activator of Nuclear Factor kB Ligand (RANKL) and von Willebrand factor (vWF) in Human Umbilical Vein Endothelial Cells (HUVEC) culture. HUVEC were isolated from human umbilical vein by a standard method. The supernatant concentrations (ELISA) and gene expression (Real Time-PCR) of OPG, RANKL and vWF in HUVEC were determined after incubation with various concentrations of UFH, enoxaparin and sulodexide for up to 16 hours. In control HUVEC strong positive correlation between OPG and vWF levels was observed, whereas sRANKL negatively correlated with OPG and vWF levels. Only in control HUVEC a negative correlation between the supernatant level of vWF and its gene expression was found. Already the lowest concentration of UFH caused 2.5-fold increase in OPG gene expression while higher UFH concentrations substantially increased RANKL mRNA level. A negative correlation between the OPG and sRANKL concentration was noticed in supernatant HUVEC which were incubated with enoxaparine. In conclusion, the observed interrelationships between OPG, RANKL and vWF levels in unstimulated HUVEC support the presumption of the pathophysiological links between these proteins. Of the tested heparin formulas UFH seems to be the most potent in altering the OPG, RANKL and vWF axis.
Keywords
heparins; HUVEC; osteoprotegerin; sRANKL; sulodexide; von Willebrand Factor
Title
Effect of unfractionated heparin, enoxaparin and sulodexide on the relations between secretion and expression of OPG, RANKL and vWF in HUVEC
Journal
Folia Histochemica et Cytobiologica
Issue
Article type
Original paper
Pages
156-163
Published online
2013-08-02
Page views
1895
Article views/downloads
3323
DOI
10.5603/FHC.2013.0016
Bibliographic record
Folia Histochem Cytobiol 2013;51(2):156-163.
Keywords
heparins
HUVEC
osteoprotegerin
sRANKL
sulodexide
von Willebrand Factor
Authors
Katarzyna Lekesiz
Beata Naumnik
Hanna Borysewicz-Sanczyk
Ewa Koc-Zurawska
Michal Mysliwiec