Farivar Alexander S, Woolley Steven M, Naidu Babu V, Fraga Charles H, Byrne Karen, Thomas Robert, Salzman Andrew L, Szabo Csaba S, Mulligan Michael S
Department of Surgery, Division of Cardiothoracic Surgery, University of Washington Medical Center, 1959 NE Pacific Street, Seattle, WA 98195, USA.
J Heart Lung Transplant. 2004 Aug;23(8):993-1002. doi: 10.1016/j.healun.2003.08.009.
Obliterative bronchiolitis (OB) is the major long-term complication affecting lung transplant recipients, and is characterized pathologically by chronic inflammatory and fibroproliferative airway disease. Based on studies revealing anti-inflammatory and anti-apoptotic properties of poly (ADP)-ribose synthetase (PARS) inhibitors, we hypothesized that their administration would be protective in a heterotopic model of experimental OB.
We transplanted rat tracheas from Brown-Norway donors into Lewis recipients, and treated 2 groups with a novel PARS inhibitor, INO-1001. One group received 14 days of treatment, whereas a second received delayed treatment beginning on Day 7 post-transplant. Tracheas were analyzed by light microscopy and computerized morphometry. Effects on cytokine transcription, nuclear transcription factor activation and cellular death were assessed by in situ hybridization for tumor necrosis factor-alpha (TNF-alpha), electromobility shift assays for nuclear factor-kappaB (NF-kappaB) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays, respectively.
PARS inhibition significantly decreased luminal obstruction (p < 0.001) and enhanced preservation of epithelial lining (p < 0.001) at 14 days post-transplant. Day 7 controls confirmed the development of an obstructive lesion in the lumen, averaging 28% occlusion. Delayed treatment (beginning on Day 7) arrested (p < 0.001) progression of the established lesion. Allograft airways treated with INO-1001 demonstrated attenuated NF-kappaB nuclear translocation, reduced transcription of TNF-alpha mRNA, and decreased cellular death on TUNEL and caspase 3 staining.
PARS inhibition is anti-inflammatory, protects against experimental OB, and is associated with enhanced preservation of respiratory epithelium and decreased cellular death. Delayed treatment with INO-1001 arrests progression of the lesion developed by Day 7. These studies suggest that activation of PARS plays a critical role in the development of airway obliterative disease.
闭塞性细支气管炎（OB）是影响肺移植受者的主要长期并发症，其病理特征为慢性炎症和纤维增生性气道疾病。基于揭示聚（ADP）-核糖合成酶（PARS）抑制剂具有抗炎和抗凋亡特性的研究，我们推测给予该抑制剂在实验性OB的异位模型中具有保护作用。
我们将来自Brown-Norway供体的大鼠气管移植到Lewis受体中，并用新型PARS抑制剂INO-1001治疗两组。一组接受14天的治疗，而另一组在移植后第7天开始接受延迟治疗。通过光学显微镜和计算机形态学分析气管。分别通过肿瘤坏死因子-α（TNF-α）的原位杂交、核因子-κB（NF-κB）的电泳迁移率变动分析和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）分析评估对细胞因子转录、核转录因子激活和细胞死亡的影响。
移植后14天，PARS抑制显著降低管腔阻塞（p < 0.001）并增强上皮内衬的保存（p < 0.001）。第7天的对照组证实管腔内出现阻塞性病变，平均阻塞率为28%。延迟治疗（从第7天开始）阻止了（p < 0.001）已形成病变的进展。用INO-1001治疗的同种异体移植气道显示NF-κB核转位减弱、TNF-α mRNA转录减少以及TUNEL和半胱天冬酶3染色显示细胞死亡减少。
PARS抑制具有抗炎作用，可预防实验性OB，并且与增强呼吸道上皮的保存和减少细胞死亡有关。用INO-1001进行延迟治疗可阻止第7天形成的病变进展。这些研究表明PARS的激活在气道闭塞性疾病的发展中起关键作用。
