Hubei Key Laboratory of Medical Technology on Transplantation, Transplant Center of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Zhongnan Hospital of Wuhan University, Wuhan, PR China.
Research Center of National Health Ministry on Transplantation Medicine Engineering and Technology, The 3rd Xiangya Hospital of Central South University, Changsha, PR China.
Artif Organs. 2021 Sep;45(9):E304-E316. doi: 10.1111/aor.13976. Epub 2021 Jun 9.
In this study, we evaluated the restoring and defatting effect of hypothermic oxygenated perfusion (HOPE) on severe steatotic liver grafts with a defatting cocktail (DF) in a rat model. Severe (≥60%) hepatic macrosteatosis was induced by a high-fat diet (HFD) for 6 weeks, after which the rats were randomly divided into four following groups: Control group, with lean livers being preserved in static cold storage (SCS) at 0°C-4°C for 45 minutes; SCS group, with a steatotic liver graft (SLG) being preserved in SCS at 0°C-4°C for 4 hours; HOPE group, where SLG was perfused with 3-hours HOPE followed by 1-hours SCS; and HOPE + DF group, HOPE with the addition of DF. Graft function after orthotopic liver transplantation was assessed in terms of mitochondrial function (adenosine triphosphate [ATP], Glycogen), endoplasmic reticulum stress (PPY, GRP78, CHOP, and ATF-6), cell apoptosis (Tunel assay, Caspase-3), inflammatory level (HMGB1, TLR4, IL-1β, IL-6. TNF-α, Factor V), and posttransplantation survival. HOPE protected steatotic liver grafts from microcirculation disturbance and endoplasmic reticulum stress and then promoted ATP and glycogen synthesis that improved mitochondrial function. Meanwhile, under conditions of ischemia-reperfusion injury, it prevented nuclear injury and endothelial damage by suppressing the release of an inflammatory mediator. The high efficacy of HOPE was enhanced after the addition of the DF. DF agents cannot promote the lipid decomposition of the steatotic liver graft at 0°C-4°C, but they can further improve steatotic liver and postoperative survival compared to the HOPE. The defatted steatotic liver grafts can be safely used in rat orthotopic liver transplantation.
在这项研究中，我们使用低温氧合灌注（HOPE）和脱脂鸡尾酒（DF）在大鼠模型中评估了低温氧合灌注对具有 DF 的重度脂肪变性肝移植物的修复和脱脂效果。通过高脂饮食（HFD）喂养 6 周诱导严重（≥60%）肝脂肪变性，然后将大鼠随机分为以下四组：对照组，将瘦肝在 0°C-4°C 的静态冷保存（SCS）中保存 45 分钟；SCS 组，将脂肪变性肝移植物（SLG）在 0°C-4°C 的 SCS 中保存 4 小时；HOPE 组，SLG 用 3 小时 HOPE 灌注，然后用 1 小时 SCS；HOPE+DF 组，HOPE 中加入 DF。通过评估线粒体功能（三磷酸腺苷 [ATP]、糖原）、内质网应激（PPY、GRP78、CHOP、ATF-6）、细胞凋亡（Tunel 检测、Caspase-3）、炎症水平（HMGB1、TLR4、IL-1β、IL-6、TNF-α、因子 V）和移植后存活率来评估原位肝移植后的移植物功能。HOPE 可保护脂肪变性肝移植物免受微循环障碍和内质网应激，并促进 ATP 和糖原合成，从而改善线粒体功能。同时，在缺血再灌注损伤的情况下，通过抑制炎症介质的释放来防止核损伤和内皮损伤。DF 的添加增强了 HOPE 的高效性。DF 制剂不能在 0°C-4°C 促进脂肪变性肝移植物的脂解，但与 HOPE 相比，它们可以进一步改善脂肪变性肝和术后存活率。脱脂脂肪变性肝移植物可安全用于大鼠原位肝移植。
