Noda Kentaro, Atale Neha, Austin Taylor, Geller David A, Gerlach Jorg, Sanchez Pablo G
Department of Cardiothoracic Surgery, University of Pittsburgh Medical Center Health System, Pittsburgh, PA, USA.
Department of Surgery, University of Pittsburgh Medical Center Health System, Pittsburgh, PA, USA.
Int J Artif Organs. 2025 Feb;48(2):84-91. doi: 10.1177/03913988251315092. Epub 2025 Jan 30.
as we look to extend lung perfusion times (EVLP) to improve preservation, the metabolic activity of the lungs will require support from other organ functions. Active functional liver support, including detoxification, synthesis, and regulation, can improve lung preservation during EVLP. This study aimed to demonstrate the effects of hepatic conditioning of the EVLP perfusate on lung endothelium, via the receptor of advanced glycation end-products (RAGE)-nuclear-factor-κB (NF-κB) signaling in vitro.
we performed in vitro experiments using human lung microvascular endothelial cells (HLMVECs), human hepatocytes, and perfusate (Steen solution). Four experimental groups: 1) fresh Steen (negative controls, NC), 2) EVLP'ed Steen control, this solution collected after 12 h of EVLP of human lungs, 3) hepatocyte conditioned EVLP'ed Steen (Hep-cond.), and 4) a RAGE inhibitor added in EVLP'ed Steen (RAGE inhibitor). HLMVECs were incubated in each testing condition and exposed to hypoxia (1% O/8% CO) for 24 h. Media were collected to investigate NF-κB signaling and endothelial glycocalyx damage.
HLMVECs incubated under hypoxia in EVLP'ed Steen showed significantly upregulated NF-κB signal and endothelial damage denoted by increased glycosaminoglycans and matrix metalloproteinase-2 activity among the groups. The Hep-cond. solution significantly attenuated those findings, while the RAGE inhibitor attenuated the NF-κB signal but not endothelial glycocalyx damage.
Our study demonstrates that hepatic function incorporated into EVLP can ameliorate pulmonary endothelial cells injury under hypoxic normothermic perfusion exposure. Our data supports the concept of incorporating other organ functions into an organ perfusion platform, to enhance lung graft preservation.
随着我们寻求延长肺灌注时间（体外肺灌注）以改善肺保存效果，肺的代谢活动将需要其他器官功能的支持。积极的功能性肝脏支持，包括解毒、合成和调节，可以改善体外肺灌注期间的肺保存。本研究旨在通过体外晚期糖基化终产物受体（RAGE）-核因子-κB（NF-κB）信号通路，证明体外肺灌注灌注液的肝脏预处理对肺内皮的影响。
我们使用人肺微血管内皮细胞（HLMVECs）、人肝细胞和灌注液（Steen溶液）进行了体外实验。四个实验组：1）新鲜Steen溶液（阴性对照，NC），2）体外肺灌注后的Steen对照溶液，该溶液在人肺体外肺灌注12小时后收集，3）肝细胞预处理的体外肺灌注Steen溶液（肝预处理），4）在体外肺灌注的Steen溶液中添加RAGE抑制剂（RAGE抑制剂）。将HLMVECs在每种测试条件下孵育，并暴露于低氧（1%O/8%CO）环境24小时。收集培养基以研究NF-κB信号通路和内皮糖萼损伤。
在体外肺灌注的Steen溶液中低氧孵育的HLMVECs显示，各组中NF-κB信号显著上调，糖胺聚糖和基质金属蛋白酶-2活性增加表明内皮损伤。肝预处理溶液显著减轻了这些结果，但RAGE抑制剂减弱了NF-κB信号，但未减轻内皮糖萼损伤。
我们的研究表明，将肝功能纳入体外肺灌注可以改善低氧常温灌注暴露下的肺内皮细胞损伤。我们的数据支持将其他器官功能纳入器官灌注平台以增强肺移植保存的概念。
