In healthy subjects fibrinogen γ/γ‘ circulates at 8–15% of the total plasma fibrinogen concentration. Elevated levels of this variant have been associated with arterial thrombosis, and its diminution with venous thrombosis. The aims of the present work were to analyze the structure of the fibrin network formed on the top of human dermal microvascular endothelial cells (HMEC-1) at different fibrinogen γ/γ‘ concentrations, as well as its influence on the secretion of fibrinolytic components. The kinetics of fibrin polymerization on top of HMEC-1 cells with 3, 10, and 30% fibrinogen γ/γ‘ was followed at 350 nm. The secretion of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type 1 (PAI 1) by HMEC-1 were measured in the supernatant and cell lysates, after incubation with 1 nM thrombin, fibrin with 3, and 30% fibrinogen γ/γ‘, using commercial kits. The influence of fibrinogen γ/γ‘ on fibrin structure on the surface of the HMEC-1 was followed with laser scanning confocal microscopy (LSCM). The kinetics of fibrin formation on HMEC-1 with 3 and 10% fibrinogen γ/γ‘ were similar. However, with 30% fibrinogen γ/γ‘ both the slope and final turbity were approximately 50% less. The LSCM images showed the dramatic effects of increasing fibrinogen γ/γ‘ from 3 to 30%. The uPA and PAI 1 concentrations in culture supernatants HMEC-1 cells treated with thrombin or 30% γ/γ‘ fibrin were two-fold increased as compared to basal culture supernatants and 3% γ/γ‘ fibrin-treated HMEC-1. In all stimulatory conditions the intracellular concentration of uPA was higher than in supernatants. In contrast, the intracellular PAI 1 concentration was decreased as compared to that measured in the supernatant, including the basal condition. A concentration of 30% fibrin γ/γ‘ alter drastically fibrin structure on the cell surface and affects the secretion of uPA and PAI 1 through its capacity to bind thrombin.

中文翻译：

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纤维蛋白γ/γ'影响纤溶成分和凝块结构的分泌

在健康受试者中，纤维蛋白原γ/γ'的循环占血浆纤维蛋白原总浓度的8-15％。该变体水平升高与动脉血栓形成有关，其减少与静脉血栓形成有关。本工作的目的是分析在不同纤维蛋白原γ/γ'浓度下在人真皮微血管内皮细胞（HMEC-1）顶部形成的纤维蛋白网络的结构，以及其对纤维蛋白溶解成分分泌的影响。 。在350 nm处跟踪具有3％，10％和30％纤维蛋白原γ/γ'的HMEC-1细胞顶部的纤维蛋白聚合动力学。在用1 nM凝血酶，3的纤维蛋白孵育后，测量上清液和细胞裂解物中HMEC-1分泌的尿激酶型纤溶酶原激活物（uPA）和纤溶酶原激活物抑制剂1（PAI 1）的分泌。和使用商业试剂盒的30％纤维蛋白原γ/γ'。激光扫描共聚焦显微镜（LSCM）追踪纤维蛋白原γ/γ'对HMEC-1表面纤维蛋白结构的影响。在含3％和10％纤维蛋白原γ/γ'的HMEC-1上形成纤维蛋白的动力学是相似的。但是，使用30％的纤维蛋白原γ/γ'时，斜率和最终湍流度都降低了约50％。LSCM图像显示出将纤维蛋白原γ/γ'从3％增加到30％的显着效果。与基础培养上清液和3％γ/γ'血纤蛋白处理过的HMEC-1相比，用凝血酶或30％γ/γ'血纤蛋白处理过的培养上清液HMEC-1细胞中的uPA和PAI 1浓度增加了两倍。在所有刺激条件下，uPA的细胞内浓度均高于上清液。相比之下，与包括基础条件在内的上清液相比，细胞内PAI 1浓度降低。浓度为30％的血纤蛋白γ/γ'会大大改变细胞表面的血纤蛋白结构，并通过其结合凝血酶的能力影响uPA和PAI 1的分泌。