Intratracheal Administration of Small Interfering RNA Targeting Fas Reduces Lung Ischemia-Reperfusion Injury
Intratracheal Administration of Small Interfering RNA Targeting Fas Reduces Lung Ischemia-Reperfusion Injury
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DOI：
10.1097/ccm.0000000000001601
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发表时间：
2016-08-01
影响因子：
8.8
通讯作者：
Ranieri, V. Marco
中科院分区：
文献类型：
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作者：
Del Sorbo, Lorenzo;Costamagna, Andrea;Ranieri, V. Marco
目的：肺缺血再灌注损伤是肺移植术后原发性移植物功能障碍的主要原因，并导致发病率和死亡率增加。Fas介导的细胞凋亡是缺血再灌注损伤的病理机制之一。我们假设，在复制肺移植程序序列的离体模型中，通过肺内施用小干扰RNA抑制肺中Fas基因表达可以减少肺缺血再灌注损伤。设计：前瞻性、随机、对照实验研究。设置：大学研究实验室。受试者：C57/BL 6小鼠，体重28-30 g。干预措施：在用靶向Fas的肺内小干扰RNA、对照小干扰RNA或载体处理后48小时，在从小鼠分离的肺中诱导缺血-再灌注损伤。离体肺暴露于6小时的冷缺血（4摄氏度），随后是2小时的温暖（37摄氏度）再灌注的溶液中含有10%的新鲜全血和机械通气与恒定的低驱动压力。缺血后Fas基因表达在mRNA和蛋白水平上显著沉默，与用对照小干扰RNA或媒介物处理的肺相比，用靶向Fas的小干扰RNA处理的肺的再灌注。Fas基因表达沉默导致水肿形成减少（支气管肺泡灌洗蛋白浓度和肺组织学）和肺顺应性改善。这些作用与靶向Fas的小干扰RNA处理的肺细胞凋亡显着减少有关，但不影响细胞因子释放和中性粒细胞浸润。结论：小干扰RNA沉默肺中Fas表达可有效对抗缺血再灌注损伤。这种方法代表了肺移植前器官保存的潜在创新策略。
Objectives: Lung ischemia-reperfusion injury is the main cause of primary graft dysfunction after lung transplantation and results in increased morbidity and mortality. Fas-mediated apoptosis is one of the pathologic mechanisms involved in the development of ischemia-reperfusion injury. We hypothesized that the inhibition of Fas gene expression in lungs by intratracheal administration of small interfering RNA could reduce lung ischemia-reperfusion injury in an ex vivo model reproducing the procedural sequence of lung transplantation.Design: Prospective, randomized, controlled experimental study.Setting: University research laboratory.Subjects: C57/BL6 mice weighing 28-30 g.Interventions: Ischemia-reperfusion injury was induced in lungs isolated from mice, 48 hours after treatment with intratracheal small interfering RNA targeting Fas, control small interfering RNA, or vehicle. Isolated lungs were exposed to 6 hours of cold ischemia (4 degrees C), followed by 2 hours of warm (37 degrees C) reperfusion with a solution containing 10% of fresh whole blood and mechanical ventilation with constant low driving pressure.Measurements and Main Results: Fas gene expression was significantly silenced at the level of messenger RNA and protein after ischemia-reperfusion in lungs treated with small interfering RNA targeting Fas compared with lungs treated with control small interfering RNA or vehicle. Silencing of Fas gene expression resulted in reduced edema formation (bronchoalveolar lavage protein concentration and lung histology) and improvement in lung compliance. These effects were associated with a significant reduction of pulmonary cell apoptosis of lungs treated with small interfering RNA targeting Fas, which did not affect cytokine release and neutrophil infiltration.Conclusions: Fas expression silencing in the lung by small interfering RNA is effective against ischemia-reperfusion injury. This approach represents a potential innovative strategy of organ preservation before lung transplantation.