Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei 430071, P.R. China.
Mol Med Rep. 2020 Sep;22(3):2003-2011. doi: 10.3892/mmr.2020.11257. Epub 2020 Jun 18.
Hypothermic machine perfusion (HMP) is a method that can be more effective in preserving donor organs compared with cold storage (CS). However, the optimal duration and the exact mechanisms of the protevtive effects of HMP remain unknow. The present study aimed to investigate the adequate perfusion time and mechanisms underlying HMP to protect livers donated after circulatory death (DCD). After circulatory death, adult male Sprague‑Dawley rat livers were subjected to 30 min of warm ischemia (WI) and were subsequently preserved by HMP or CS. To determine the optimal perfusion time, liver tissues were analyzed at 0, 1, 3, 5, 12 and 24 h post‑preservation to evaluate injury and assess the expression of relevant proteins. WI livers were preserved by HMP or CS for 3 h, and liver viability was evaluated by normothermic reperfusion (NR). During NR, oxygen consumption, bile production and the activities of hepatic enzymes in the perfusate were assessed. Following 2 h of NR, levels of inflammation and oxidative stress were determined in the livers and perfusate. HMP for 3 h resulted in the highest expression of myocyte enhancer factor 2C (MEF2C) and kruppel‑like factor 2 (KLF2) and the lowest expression of NF‑κB p65, tumor necrosis factor (TNF)‑α and interleukin (IL)‑1β among the different timepoints, which indicated that 3 h may be the optimal time for HMP induction of the KLF2‑dependent signaling pathway. Compared with CS‑preserved livers, HMP‑preserved livers displayed significantly higher oxygen consumption, lower hepatic enzyme levels in the perfusate following NR. Following HMP preservation, the expression levels of MEF2C, KLF2, endothelial nitric oxide synthase and nitric oxide were increased, whereas the expression levels of NF‑κB p65, IL‑1β and TNF‑α were decreased compared with CS preservation. The results indicated that 3 h may be the optimal time for HMP to protect DCD rat livers. Furthermore, HMP may significantly reduce liver inflammation and oxidative stress injury by mediating the KLF2/NF‑κB/eNOS‑dependent signaling pathway.
低温机器灌注（HMP）是一种与低温保存（CS）相比可以更有效地保存供体器官的方法。然而，HMP 的最佳灌注时间和确切保护作用机制仍不清楚。本研究旨在探讨 HMP 保护心跳骤停后供体肝脏的适当灌注时间和机制。心跳骤停后，成年雄性 Sprague-Dawley 大鼠肝脏经历 30 分钟的热缺血（WI），随后通过 HMP 或 CS 保存。为了确定最佳灌注时间，在保存后 0、1、3、5、12 和 24 小时分析肝组织，以评估损伤并评估相关蛋白的表达。WI 肝脏通过 HMP 或 CS 保存 3 小时，通过常温再灌注（NR）评估肝存活率。在 NR 期间，评估灌流液中的耗氧量、胆汁生成和肝酶活性。NR 2 小时后，测定肝脏和灌流液中的炎症和氧化应激水平。NR 期间，测定肝脏和灌流液中的炎症和氧化应激水平。HMP 灌注 3 小时后，肌细胞增强因子 2C（MEF2C）和 Kruppel 样因子 2（KLF2）的表达最高，NF-κB p65、肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β的表达最低，这表明 3 小时可能是 HMP 诱导 KLF2 依赖性信号通路的最佳时间。与 CS 保存的肝脏相比，HMP 保存的肝脏在 NR 后灌流液中的耗氧量更高，肝酶水平更低。HMP 保存后，MEF2C、KLF2、内皮型一氧化氮合酶和一氧化氮的表达水平增加，而 NF-κB p65、IL-1β和 TNF-α的表达水平降低。结果表明，HMP 保护 DCD 大鼠肝脏的最佳时间可能为 3 小时。此外，HMP 可能通过调节 KLF2/NF-κB/eNOS 依赖性信号通路，显著减轻肝脏炎症和氧化应激损伤。
