Lekesiz Katarzyna, Naumnik Beata, Borysewicz-Sanczyk Hanna, Koc-Zurawska Ewa, Mysliwiec Michal
Department of Nephrology and Transplantation with Dialysis Unit, Medical University, Bialystok, Poland.
Folia Histochem Cytobiol. 2013;51(2):156-63. doi: 10.5603/FHC.2013.0016.
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.
肝素可调节血管内皮功能。我们研究了普通肝素（UFH）、依诺肝素和舒洛地昔对人脐静脉内皮细胞（HUVEC）培养物中骨保护素（OPG）、核因子κB受体活化因子配体（RANKL）和血管性血友病因子（vWF）水平及基因表达的影响。通过标准方法从人脐静脉分离出HUVEC。在用不同浓度的UFH、依诺肝素和舒洛地昔孵育长达16小时后，测定HUVEC中OPG、RANKL和vWF的上清液浓度（酶联免疫吸附测定法）和基因表达（实时聚合酶链反应）。在对照HUVEC中，观察到OPG与vWF水平之间呈强正相关，而可溶性RANKL（sRANKL）与OPG和vWF水平呈负相关。仅在对照HUVEC中发现vWF上清液水平与其基因表达之间呈负相关。即使是最低浓度的UFH也会使OPG基因表达增加2.5倍，而较高浓度的UFH会显著增加RANKL mRNA水平。在用依诺肝素孵育的HUVEC上清液中，观察到OPG与sRANKL浓度之间呈负相关。总之，在未受刺激的HUVEC中观察到的OPG、RANKL和vWF水平之间的相互关系支持了这些蛋白质之间存在病理生理联系的推测。在所测试的肝素配方中，UFH似乎在改变OPG、RANKL和vWF轴方面最有效。
