Manipulation of Lipid Metabolism During Normothermic Machine Perfusion: Effect of Defatting Therapies on Donor Liver Functional Recovery
Manipulation of Lipid Metabolism During Normothermic Machine Perfusion: Effect of Defatting Therapies on Donor Liver Functional Recovery
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DOI：
10.1002/lt.25439
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发表时间：
2019-07-01
影响因子：
4.6
通讯作者：
Afford, Simon C.
中科院分区：
文献类型：
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作者：
Boteon, Yuri L.;Attard, Joseph;Afford, Simon C.
需要增加脂肪变性供体肝脏使用的策略来解决移植等待名单上的死亡率。我们的目的是测试在离体常温机器灌注过程中药理学增强人肝脏脂质代谢以促进脱脂和改善器官功能恢复的功效。由于脂肪变性，丢弃10个肝脏，并分配到一个脱脂组，该组的灌注液补充了一种药物组合，以提高脂质代谢，或对照组，接受灌注液与车辆。使用组织匀浆和组织学分析评估脂肪变性。通过酶联免疫吸附试验、免疫组织化学和凝胶内蛋白检测评价脂质氧化和溶解、氧化损伤、炎症和胆道功能的标志物。在6小时内，治疗使组织甘油三酯减少38%，大泡性脂肪变性减少40%。这种效应是由甘油三酯溶解度增加（P = 0.04）驱动的，通过增加生酮（P = 0.008）和三磷酸腺苷合成（P = 0.01）评估的线粒体氧化与酰基辅酶A氧化酶1、肉毒碱棕榈酰转移酶1A和乙酰辅酶A合成酶水平增加相关。同时，脱脂肝脏表现出代谢功能参数增强，如尿素产生（P = 0.03），血管阻力降低，丙氨酸氨基转移酶释放降低（P = 0.049），胆汁产生增加（P = 0.008），胆汁pH值升高（P = 0.03）。治疗下调了氧化损伤标志物的表达以及免疫细胞的活化（CD 14; CD 11b），并减少了灌注液中炎性细胞因子（肿瘤坏死因子α;白细胞介素1 β）的释放。总之，在常温机器灌注过程中，药理学增强细胞内脂质代谢可在6小时内降低人肝脏的脂质含量。它还改善了对器官的细胞内代谢支持，导致成功的功能恢复和再灌注损伤标志物的表达降低。
Strategies to increase the use of steatotic donor livers are required to tackle the mortality on the transplant waiting list. We aimed to test the efficacy of pharmacological enhancement of the lipid metabolism of human livers during ex situ normothermic machine perfusion to promote defatting and improve the functional recovery of the organs. Because of steatosis, 10 livers were discarded and were allocated either to a defatting group that had the perfusate supplemented with a combination of drugs to enhance lipid metabolism or to a control group that received perfusion fluid with vehicle only. Steatosis was assessed using tissue homogenate and histological analyses. Markers for lipid oxidation and solubilization, oxidative injury, inflammation, and biliary function were evaluated by enzyme-linked immunosorbent assay, immunohistochemistry, and in-gel protein detection. Treatment reduced tissue triglycerides by 38% and macrovesicular steatosis by 40% over 6 hours. This effect was driven by increased solubility of the triglycerides (P = 0.04), and mitochondrial oxidation as assessed by increased ketogenesis (P = 0.008) and adenosine triphosphate synthesis (P = 0.01) were associated with increased levels of the enzymes acyl-coenzyme A oxidase 1, carnitine palmitoyltransferase 1A, and acetyl-coenzyme A synthetase. Concomitantly, defatted livers exhibited enhanced metabolic functional parameters such as urea production (P = 0.03), lower vascular resistance, lower release of alanine aminotransferase (P = 0.049), and higher bile production (P = 0.008) with a higher bile pH (P = 0.03). The treatment down-regulated the expression of markers for oxidative injury as well as activation of immune cells (CD14; CD11b) and reduced the release of inflammatory cytokines in the perfusate (tumor necrosis factor alpha; interleukin 1 beta). In conclusion, pharmacological enhancement of intracellular lipid metabolism during normothermic machine perfusion decreased the lipid content of human livers within 6 hours. It also improved the intracellular metabolic support to the organs, leading to successful functional recovery and decreased expression of markers of reperfusion injury.