Biedermann B, Rosenmund A, Muller M, Kohler H P, Haeberli A, Straub P W
Department of Medicine, University Hospital of Bern, Switzerland.
J Lab Clin Med. 1994 Sep;124(3):339-47.
Human endothelial cells cultivated on polystyrene microcarrier beads were used to study endothelial anticoagulant activity in vitro. Spontaneous whole blood coagulation was inhibited by endothelial cells on microcarriers at a surface to volume ratio of 16 cm2/ml blood. Thrombin activity generated in nonanticoagulated whole blood during 1 hour and assessed by its fibrinogen clotting effect was reduced by 87% in the presence of endothelial cells. Consistent with this observation, prothrombin fragment1+2, fibrinopeptide A, and thrombin-antithrombin III-complex release during the same period of time were inhibited by 81%, 47%, and 88%, respectively. Immunoblotting analysis of cell-free supernatants derived from the same samples demonstrated that prothrombin activation was strongly suppressed in the presence of endothelial cells. Furthermore, the incubation of nonanticoagulated whole blood with endothelialized beads for only 5 minutes after venipuncture was sufficient to prevent subsequent prothrombin activation in the cell-free supernatants of the same whole blood sample after centrifugation. These findings suggest that interruption of the coagulation cascade is probably one major mechanism of endothelial anticoagulant activity in vivo.
将在聚苯乙烯微载体珠上培养的人内皮细胞用于体外研究内皮抗凝活性。微载体上的内皮细胞以16 cm²/ml血液的表面积与体积比抑制自发全血凝固。通过纤维蛋白原凝血作用评估，在存在内皮细胞的情况下，未抗凝全血在1小时内产生的凝血酶活性降低了87%。与此观察结果一致，同期凝血酶原片段1+2、纤维蛋白肽A和凝血酶-抗凝血酶III复合物的释放分别被抑制了81%、47%和88%。对来自相同样本的无细胞上清液进行免疫印迹分析表明，在存在内皮细胞的情况下，凝血酶原激活受到强烈抑制。此外，静脉穿刺后仅用内皮化珠孵育未抗凝全血5分钟，就足以防止同一全血样本离心后的无细胞上清液中随后的凝血酶原激活。这些发现表明，凝血级联反应的中断可能是体内内皮抗凝活性的一种主要机制。
