Hypothermic oxygenated machine perfusion alleviates liver injury in donation after circulatory death through activating autophagy in mice
Hypothermic oxygenated machine perfusion alleviates liver injury in donation after circulatory death through activating autophagy in mice
复制标题
低温充氧机灌注通过激活小鼠自噬减轻循环死亡后捐献肝损伤
DOI：
10.1111/aor.13525
复制
发表时间：
2019-07-18
影响因子：
2.4
通讯作者：
Ye, Qifa
中科院分区：
文献类型：
--
作者：
Zeng, Xianpeng;Wang, Shengjie;Ye, Qifa
低温氧合机灌注（HOPE）是一种安全可靠的方法，可以减轻循环死亡后捐献（DCD）的肝损伤。本研究重点探讨自噬在 HOPE 对 DCD 肝损伤的保护作用中的作用。建立小鼠30分钟温缺血肝模型。冷藏（CS）4小时后，采用100％O-2或100％N-2低温机器灌注（HMP）1小时。再灌注2小时内，收集肝组织和灌注液，评价肝功能、氧化应激水平、细胞凋亡和坏死。使用蛋白质印迹法探讨自噬水平。当肝脏经历热缺血损伤时，LC3B-II表达显着增强。与CS相比，HOPE诱导较低的AST和ALT释放，以及较低的氧化应激水平、细胞凋亡和坏死细胞数量，并导致组织ATP含量较高。与此同时，自噬相关蛋白（如 ULK1、Atg5 和 LC3B-II）的表达增加。当氧气完全被氮气取代时，HMP的清除作用并没有激活自噬，也不能缓解DCD肝损伤。当HOPE中使用自噬抑制剂3-甲基腺嘌呤时，HOPE的保护作用减弱。总之，与健康肝脏相比，DCD 肝损伤激活了自噬，而 HOPE 通过进一步增加该小鼠模型中的自噬水平来减轻 DCD 肝损伤。然而，与 CS 相比，含有 100% N-2 的 HMP 对 DCD 肝损伤或自噬水平没有有益作用。自噬的研究可能为临床减轻DCD肝损伤提供新策略。
Hypothermic oxygenated machine perfusion (HOPE) is a safe and reliable method that could alleviate liver injury in donation after circulatory death (DCD). This study focuses on the role of autophagy in HOPE's protective effect on DCD liver injury. A 30-minute warm ischemic liver model was established in mice. After 4 hours of cold storage (CS), 1 hour of hypothermic machine perfusion (HMP) with 100% O-2 or 100% N-2 was employed. During 2 hours of reperfusion, liver tissue and perfusate were collected to evaluate liver function, oxidative stress level, apoptosis, and necrosis. Western blotting was used to explore the level of autophagy. When the liver experienced warm ischemic injury, LC3B-II expression was significantly enhanced. Compared with the CS, HOPE induced lower release of AST and ALT, as well as lower oxidative stress levels, apoptosis, and necrosis cell numbers, and led to higher tissue ATP content. Meanwhile, expression of autophagy-related proteins, such as ULK1, Atg5, and LC3B-II, increased. When oxygen was completely replaced by nitrogen, the washout effect of HMP did not activate autophagy and did not relieve DCD liver injury. When the autophagy inhibitor 3-methyladenine was used in HOPE, the protective effect of HOPE was attenuated. In conclusion, DCD liver injury activated autophagy compared with healthy liver, while HOPE alleviated DCD liver injury by increasing autophagy levels further in this mouse model. However, HMP with 100% of N-2 had no beneficial effect on DCD liver injury or on autophagy levels compared with CS. The research on autophagy may provide a new strategy for alleviating DCD liver injury in clinical practice.