Bach-Gansmo E T, Halvorsen S, Godal H C, Skjonsberg O H
Department of Pulmonary Medicine, Ulleval Hospital, University of Oslo, Norway.
Thromb Res. 1996 Apr 15;82(2):177-86. doi: 10.1016/0049-3848(96)00064-3.
To see if D-dimers were degraded by human neutrophil elastase (HNE), cross-linked fibrin was obtained by adding thrombin to purified fibrinogen in the presence of calcium ions and factor XIII, and the fibrin clot subsequently degraded by plasmin. Thereafter, the supernatant containing fibrin degradation products was removed and incubated with HNE. D-dimer levels were measured by two rapid semiquantitative tests, a latex agglutination test and the Nycocard immunofiltration test, and a quantitative ELISA-method. With increasing incubation time, D-dimer levels as measured by the latex and Nycocard tests rapidly decreased and subsequently became undetectable, while the ELISA D-dimer values remained essentially unchanged. By using SDS-electrophoresis and immunoblotting, the degradation of plasmic derivatives of cross-linked fibrin by fiNE was visualised. We conclude that in a purified system, D-dimers formed during plasmin mediated lysis of cross linked fibrin are further degraded by HNE. Such HNE degradation reduces the D-dimer concentration as measured by rapid semiquantitive tests, and may be partly responsible for discrepant results when using different D-dimer assays.
为了观察D - 二聚体是否会被人中性粒细胞弹性蛋白酶（HNE）降解，在钙离子和因子XIII存在的情况下，通过向纯化的纤维蛋白原中加入凝血酶来获得交联纤维蛋白，随后纤维蛋白凝块被纤溶酶降解。此后，去除含有纤维蛋白降解产物的上清液，并与HNE一起孵育。通过两种快速半定量试验（乳胶凝集试验和Nycocard免疫过滤试验）以及定量ELISA方法测量D - 二聚体水平。随着孵育时间的增加，通过乳胶和Nycocard试验测量的D - 二聚体水平迅速下降，随后变得无法检测到，而ELISA法测定的D - 二聚体值基本保持不变。通过使用SDS - 电泳和免疫印迹，观察到了HNE对交联纤维蛋白的血浆衍生物的降解作用。我们得出结论，在纯化系统中，纤溶酶介导的交联纤维蛋白溶解过程中形成的D - 二聚体可被HNE进一步降解。这种HNE降解会降低通过快速半定量试验测量的D - 二聚体浓度，并且可能部分导致使用不同D - 二聚体检测方法时出现结果差异。
