Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor-1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits
Intrapleural Adenoviral Delivery of Human Plasminogen Activator Inhibitor-1 Exacerbates Tetracycline-Induced Pleural Injury in Rabbits
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DOI：
10.1165/rcmb.2012-0183oc
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发表时间：
2013-01-01
影响因子：
6.4
通讯作者：
Idell, Steven
中科院分区：
文献类型：
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作者：
Karandashova, Sophia;Florova, Galina;Idell, Steven
纤溶酶原激活物抑制物-1(PAI-1)浓度升高与胸膜损伤有关，但其对胸膜组织的影响尚不清楚。发展了一种腺病毒介导的目的基因(在巨细胞病毒启动子下表达)到兔胸膜的方法，并与LacZ和人(H)PAI-1一起使用。通过组织学、β-半乳糖苷酶染色、Western blotting、胸腔积液、灌洗液和胸膜间皮细胞的酶和免疫组织化学分析来评价转导的效率和效果。转导是选择性的，且仅限于胸膜间皮单分子层。两个基因在胸腔内的表达都是短暂的，在4~5天达到表达高峰。在第5天，HPAI-1(灌洗液中活性HPAI-1和总HPAI-1分别为40-80和200-400 NM)没有引起明显的胸膜损伤、积液或纤维化。腺病毒介导的hPAI-1转导与随后的四环素诱导的胸膜损伤导致第5天观察到的胸膜纤维化显著加重(与赋形剂和腺病毒对照样品相比，分别为P=0.029和P=0.021)。纤溶酶原激活剂胸腔内溶栓治疗(IPFT)对HPAI-1过度表达的动物和单纯四环素损伤的对照组动物均有效。胸腔内活性PAI-1的增加(从对照动物的10-15 nM增加到HPAI-1高表达动物的2040 nM)导致体内PAI-1/纤溶酶原激活物复合体的形成增加。在IPFT后10到40分钟观察到的胸腔内纤溶酶原激活活性的降低与活性PAI-1的初始浓度呈线性相关。因此，PFS中活化的PAI-1影响IPFT的预后，可能既是胸膜损伤的生物标志物，也是其治疗的分子靶点。
Elevated concentrations of plasminogen activator inhibitor-1 (PAI-1) are associated with pleural injury, but its effects on pleural organization remain unclear. A method of adenovirus-mediated delivery of genes of interest (expressed under a cytomegalovirus promoter) to rabbit pleura was developed and used with lacZ and human (h) PAI-1. Histology, beta-galactosidase staining, Western blotting, enzymatic and immunohistochemical analyses of pleural fluids (PFs), lavages, and pleural mesothelial cells were used to evaluate the efficiency and effects of transduction. Transduction was selective and limited to the pleural mesothelial monolayer. The intrapleural expression of both genes was transient, with their peak expression at 4 to 5 days. On Day 5, hPAI-1 (40-80 and 200-400 nM of active and total hPAI-1 in lavages, respectively) caused no overt pleural injury, effusions, or fibrosis. The adenovirus-mediated delivery of hPAI-1 with subsequent tetracycline-induced pleural injury resulted in a significant exacerbation of the pleural fibrosis observed on Day 5 (P = 0.029 and P = 0.021 versus vehicle and adenoviral control samples, respectively). Intrapleural fibrinolytic therapy (IPFT) with plasminogen activators was effective in both animals overexpressing hPAI-1 and control animals with tetracycline injury alone. An increase in intrapleural active PAI-1 (from 10-15 nM in control animals to 2040 nM in hPAI-1-overexpressing animals) resulted in the increased formation of PAI-1/plasminogen activator complexes in vivo. The decrease in intrapleural plasminogen-activating activity observed at 10 to 40 minutes after IPFT correlates linearly with the initial concentration of active PAI-1. Therefore, active PAI-1 in PFs affects the outcome of IPFT, and may be both a biomarker of pleural injury and a molecular target for its treatment.