Gálvez A, Gómez-Ortiz G, Díaz-Ricart M, Escolar G, González-Sarmiento R, Zurbano M J, Ordinas A, Castillo R
Servicio de Hemoterapia y Hemostasia, Hospital Clinic, Facultad de Medicina, Barcelona, Spain.
Thromb Haemost. 1997 May;77(5):975-80.
The effect of desmopressin (DDAVP) on thrombogenicity, expression of tissue factor and procoagulant activity (PCA) of extracellular matrix (ECM) generated by human umbilical vein endothelial cells cultures (HUVEC), was studied under different experimental conditions. HUVEC were incubated with DDAVP (1, 5 and 30 ng/ml) and then detached from their ECM. The reactivity towards platelets of this ECM was tested in a perfusion system. Coverslips covered with DDAVP-treated ECMS were inserted in a parallel-plate chamber and exposed to normal blood anticoagulated with low molecular weight heparin (Fragmin, 20 U/ml). Perfusions with run for 5 min at a shear rate of 800 s-1. Deposition of platelets on ECMs was significantly increased with respect to control ECMs when DDAVP was used at 5 and 30 ng/ml (p < 0.05 and p. < 0.01 respectively). The increase in platelet deposition was prevented by incubation of ECMs with an antibody against human tissue factor prior to perfusion. Immunofluorescence studies positively detected tissue factor antigen on DDAVP derived ECMs. A chromogenic assay performed under standardized conditions revealed a statistically significant increase in the procoagulant activity of the ECMs produced by ECs incubated with 30 ng/ml DDAVP (p < 0.01 vs. control samples). Northern blot analysis revealed increased levels of tissue factor mRNA in extracts from ECs exposed to DDAVP. Our data indicate that DDAVP in vitro enhances platelet adhesion to the ECMs through increased expression of tissue factor. A similar increase in the expression of tissue factor might contribute to the in vivo hemostatic effect of DDAVP.
在不同实验条件下，研究了去氨加压素（DDAVP）对人脐静脉内皮细胞培养物（HUVEC）产生的细胞外基质（ECM）的血栓形成性、组织因子表达和促凝血活性（PCA）的影响。将HUVEC与DDAVP（1、5和30 ng/ml）孵育，然后从其ECM上分离。在灌注系统中测试该ECM对血小板的反应性。将覆盖有DDAVP处理的ECM的盖玻片插入平行板腔室中，并暴露于用低分子量肝素（速碧林，20 U/ml）抗凝的正常血液中。以800 s-1的剪切速率进行5分钟的灌注。当使用5和30 ng/ml的DDAVP时，与对照ECM相比，ECM上血小板的沉积显著增加（分别为p < 0.05和p < 0.01）。在灌注前用抗人组织因子抗体孵育ECM可防止血小板沉积增加。免疫荧光研究在DDAVP衍生的ECM上阳性检测到组织因子抗原。在标准化条件下进行的显色测定显示，与30 ng/ml DDAVP孵育的EC产生的ECM的促凝血活性有统计学显著增加（与对照样品相比，p < 0.01）。Northern印迹分析显示，暴露于DDAVP的EC提取物中组织因子mRNA水平升高。我们的数据表明，DDAVP在体外通过增加组织因子的表达增强血小板对ECM的粘附。组织因子表达的类似增加可能有助于DDAVP的体内止血作用。
