Francioli Cyril, Wang Xingyu, Parapanov Roumen, Abdelnour Etienne, Lugrin Jérôme, Gronchi Fabrizio, Perentes Jean, Eckert Philippe, Ris Hans-Beat, Piquilloud Lise, Krueger Thorsten, Liaudet Lucas
Service of Thoracic Surgery, University Hospital Medical Center and Faculty of Biology and Medicine, Lausanne, Switzerland.
Service of Adult Intensive Care Medicine, University Hospital Medical Center and Faculty of Biology and Medicine, Lausanne, Switzerland.
PLoS One. 2017 Mar 21;12(3):e0173916. doi: 10.1371/journal.pone.0173916. eCollection 2017.
Damaged lung grafts obtained after circulatory death (DCD lungs) and warm ischemia may be at high risk of reperfusion injury after transplantation. Such lungs could be pharmacologically reconditioned using ex-vivo lung perfusion (EVLP). Since acute inflammation related to the activation of nuclear factor kappaB (NF-κB) is instrumental in lung reperfusion injury, we hypothesized that DCD lungs might be treated during EVLP by pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-κB. Rat lungs exposed to 1h warm ischemia and 2 h cold ischemia were subjected to EVLP during 4h, in absence (CTRL group, N = 6) or in presence of PDTC (2.5g/L, PDTC group, N = 6). Static pulmonary compliance (SPC), peak airway pressure (PAWP), pulmonary vascular resistance (PVR), and oxygenation capacity were determined during EVLP. After EVLP, we measured the weight gain of the heart-lung block (edema), and the concentration of LDH (cell damage), proteins (permeability edema) and of the cytokines IL-6, TNF-α and CINC-1 in bronchoalveolar lavage (BAL), and we evaluated NF-κB activation by the degree of phosphorylation and degradation of its inhibitor IκBα in lung tissue. In CTRL, we found significant NF-κB activation, lung edema, and a massive release of LDH, proteins and cytokines. SPC significantly decreased, PAWP and PVR increased, while oxygenation tended to decrease. Treatment with PDTC during EVLP inhibited NF-κB activation, did not influence LDH release, but markedly reduced lung edema and protein concentration in BAL, suppressed TNFα and IL-6 release, and abrogated the changes in SPC, PAWP and PVR, with unchanged oxygenation. In conclusion, suppression of innate immune activation during EVLP using the NF-κB inhibitor PDTC promotes significant improvement of damaged rat DCD lungs. Future studies will determine if such rehabilitated lungs are suitable for in vivo transplantation.
循环死亡后获得的受损肺移植（DCD肺）以及热缺血可能会使移植后发生再灌注损伤的风险升高。此类肺脏可通过体外肺灌注（EVLP）进行药物预处理。由于与核因子κB（NF-κB）激活相关的急性炎症在肺再灌注损伤中起作用，我们推测DCD肺在EVLP期间可用NF-κB抑制剂吡咯烷二硫代氨基甲酸盐（PDTC）进行治疗。将大鼠肺暴露于1小时热缺血和2小时冷缺血后，在无（对照组，N = 6）或有PDTC（2.5g/L，PDTC组，N = 6）的情况下进行4小时的EVLP。在EVLP期间测定静态肺顺应性（SPC）、气道峰压（PAWP）、肺血管阻力（PVR）和氧合能力。EVLP后，我们测量心肺阻滞的重量增加（水肿）、乳酸脱氢酶（LDH）浓度（细胞损伤）、蛋白质（通透性水肿）以及支气管肺泡灌洗（BAL）中细胞因子IL-6、TNF-α和CINC-1的浓度，并通过其抑制剂IκBα在肺组织中的磷酸化和降解程度评估NF-κB的激活情况。在对照组中，我们发现NF-κB显著激活、肺水肿以及LDH、蛋白质和细胞因子的大量释放。SPC显著降低, PAWP和PVR升高，而氧合趋于下降。EVLP期间用PDTC治疗可抑制NF-κB激活，不影响LDH释放，但显著减轻肺水肿和BAL中的蛋白质浓度，抑制TNFα和IL-6释放，并消除SPC、PAWP和PVR的变化，氧合无变化。总之，使用NF-κB抑制剂PDTC在EVLP期间抑制先天性免疫激活可显著改善受损大鼠DCD肺。未来的研究将确定此类修复后的肺是否适合体内移植。
