Targeting plasminogen activator inhibitor-1 in tetracycline-induced pleural injury in rabbits
Targeting plasminogen activator inhibitor-1 in tetracycline-induced pleural injury in rabbits
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DOI：
10.1152/ajplung.00579.2016
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发表时间：
2018-01-01
影响因子：
4.9
通讯作者：
Komissarov, Andrey A.
中科院分区：
文献类型：
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作者：
Florova, Galina;Azghani, Ali O.;Komissarov, Andrey A.
活性纤溶酶原激活物抑制物-1（派-1）升高对四环素诱导的家兔胸膜损伤的胸膜内纤溶治疗（IPFT）的结果有不利影响。为了用尿激酶原（scuPA）增强IPFT，测试了两种机制上不同的靶向派-1的方法：减缓其与尿激酶（uPA）的反应和单克隆抗体（mAb）介导的派-1失活。去除uPA的“派-1对接位点”（（179）RHRGGS（184）->（179）AAAAAA（184））处的带正电荷的残基导致派-1的抑制速率降低60倍。突变型尿激酶原（0.0625-0.5 mg/kg; n = 12）显示出与野生型scuPA相当的疗效，且未改变IPFT结局（P > 0.05）。值得注意的是，突变型uPA的派-1非依赖性胸膜内失活率比野生型酶高2倍（P < 0.05）。将派-1捕获在具有Ser（195）Ala取代的无催化活性的双链尿激酶（S195 A-tcuPA; 0.1和0.5 mg/kg）的“分子夹心”型复合物中并没有改善用scuPA（0.0625-0.5 mg/kg; n = 11）进行IPFT的功效。IPFT在MA-56 A7 C10（0.5 mg/kg; n = 2）存在下失败，MA-56 A7 C10形成稳定的胸膜内分子夹心复合物，通过阻断其向潜伏形式的转变而允许活性派-1积聚。相比之下，通过加速由mAb MA-33 B8（0.5mg/kg; n = 2）介导的活性至潜伏性转变来灭活派-1改善了用scuPA（0.25mg/kg）进行IPFT的功效。因此，在缓慢（4-8小时）的条件下，在四环素诱导的家兔胸膜损伤的纤维蛋白溶解，只有派-1的失活，但不减少其与uPA的反应速率，增强IPFT。因此，纤溶速率在不同的病理状态下不同，可能会影响派-1抑制剂的选择，以加强纤溶治疗。
Elevated active plasminogen activator inhibitor-1 (PAI-1) has an adverse effect on the outcomes of intrapleural fibrinolytic therapy (IPFT) in tetracycline-induced pleural injury in rabbits. To enhance IPFT with prourokinase (scuPA), two mechanistically distinct approaches to targeting PAI-1 were tested: slowing its reaction with urokinase (uPA) and monoclonal antibody (mAb)-mediated PAI-1 inactivation. Removing positively charged residues at the "PAI-1 docking site" ((179)RHRGGS(184)->(179)AAAAAA(184)) of uPA results in a 60-fold decrease in the rate of inhibition by PAI-1. Mutant prourokinase (0.0625-0.5 mg/kg; n = 12) showed efficacy comparable to wild-type scuPA and did not change IPFT outcomes (P > 0.05). Notably, the rate of PAI-1-independent intrapleural inactivation of mutant uPA was 2 times higher (P < 0.05) than that of the wild-type enzyme. Trapping PAI-1 in a "molecular sandwich"-type complex with catalytically inactive two-chain urokinase with Ser(195)Ala substitution (S195A-tcuPA; 0.1 and 0.5 mg/kg) did not improve the efficacy of IPFT with scuPA (0.0625-0.5 mg/kg; n = 11). IPFT failed in the presence of MA-56A7C10 (0.5 mg/kg; n = 2), which forms a stable intrapleural molecular sandwich complex, allowing active PAI-1 to accumulate by blocking its transition to a latent form. In contrast, inactivation of PAI-1 by accelerating the active-to-latent transition mediated by mAb MA-33B8 (0.5 mg/kg; n = 2) improved the efficacy of IPFT with scuPA (0.25 mg/kg). Thus, under conditions of slow (4-8 h) fibrinolysis in tetracycline-induced pleural injury in rabbits, only the inactivation of PAI-1, but not a decrease in the rate of its reaction with uPA, enhances IPFT. Therefore the rate of fibrinolysis, which varies in different pathologic states, could affect the selection of PAI-1 inhibitors to enhance fibrinolytic therapy.