Highly stable plasminogen activator inhibitor type one (VLHL PAI-1) protects fibrin clots from tissue plasminogen activator-mediated fibrinolysis.
Highly stable plasminogen activator inhibitor type one (VLHL PAI-1) protects fibrin clots from tissue plasminogen activator-mediated fibrinolysis.
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DOI：
10.3892/ijmm.20.5.683
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发表时间：
2007-11
影响因子：
5.4
通讯作者：
J. Jankun;Ansari M. Aleem;S. Selman;E. Skrzypczak‐Jankun;W. Lysiak-Szydlowska;Nicholas Grafos;H. J. Fryer;R. Greenfield
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文献类型：
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作者：
J. Jankun;Ansari M. Aleem;S. Selman;E. Skrzypczak‐Jankun;W. Lysiak-Szydlowska;Nicholas Grafos;H. J. Fryer;R. Greenfield
纤溶酶原激活剂抑制剂-1 (PAI-1) 是组织型纤溶酶原激活剂 (tPA) 的主要特异性抑制剂，tPA 通过激活纤溶酶原介导纤维蛋白凝块溶解。野生型 PAI-1 (wPAI-1) 迅速转化为潜在形式（半衰期约为 2 小时）并失去抑制 tPA 的能力。我们开发了一种半衰期非常长的 PAI-1 (VLHL PAI-1)，这是一种重组蛋白，与 wPAI-1 相比，半衰期 >700 小时。在这项研究中，评估了 VLHL PAI-1 体外抑制血栓溶解的能力。通过添加组织因子或人重组FVIIa，在补充有tPA的正常血浆中引发凝块形成。在微量滴定板读数器中通过比浊法监测的凝块溶解时间是在不同浓度的 wPAI-1 和 VLHL PAI-1 下测定的。 wPAI-1 和 VLHL PAI-1 均导致血栓溶解时间显着增加，尽管后者在较低浓度下效果稍差。 VLHL PAI-1（而非 wPAI-1）在 37 度预温育过夜后保持其抗纤维蛋白溶解活性。这些研究表明 VLHL PAI-1 是纤维蛋白凝块降解的有效抑制剂。由于 VLHL PAI-1 与 wPAI-1 相比具有较高的稳定性，这种新型 tPA 介导的纤维蛋白溶解抑制剂在直接使用或与促凝血重组 FVIIa 联合使用时可用于治疗手术和创伤患者。
Plasminogen activator inhibitor-1 (PAI-1) is the major specific inhibitor of tissue-type plasminogen activator (tPA) which mediates fibrin clot lysis through activation of plasminogen. Wild-type-PAI-1 (wPAI-1) is rapidly converted to the latent form (half-life of approximately 2 h) and loses its ability to inhibit tPA. We developed a very long half-life PAI-1 (VLHL PAI-1), a recombinant protein with a half-life >700 h compared with wPAI-1. In this study, VLHL PAI-1 was assessed for its ability to inhibit clot lysis in vitro. Clot formation was initiated in normal plasma supplemented with tPA by the addition of either tissue factor or human recombinant FVIIa. Clot lysis time, monitored turbidimetrically in a microtiter plate reader, was determined at various concentrations of wPAI-1 and VLHL PAI-1. Both wPAI-1 and VLHL PAI-1 caused a significant increase in clot lysis time, although the latter was somewhat less effective at lower concentrations. The VLHL PAI-1, but not wPAI-1, maintained its anti-fibrinolytic activity after preincubation overnight at 37 degrees. These studies demonstrate that VLHL PAI-1 is an effective inhibitor of fibrin clot degradation. Due to the high stability of VLHL PAI-1 compared with wPAI-1, this novel inhibitor of tPA-mediated fibrinolysis may have therapeutic applications for treating surgical and trauma patients when used directly or in conjunction with the procoagulant recombinant FVIIa.