Department of General, Visceral- and Transplantation Surgery, Hannover Medical School, Hannover, Germany.
Centre for Pharmacology and Toxicology, Hannover Medical School, Hannover, Germany.
Transl Res. 2024 Feb;264:33-65. doi: 10.1016/j.trsl.2023.09.006. Epub 2023 Sep 17.
Orthotopic liver transplantation (OLT) is a lifesaving procedure. However, grafts may fail due to primary nonfunction (PNF). In the past, we demonstrated PNFs to be mainly associated with fatty allografts, and given its unpredictable nature, the development of a disease model is urgently needed. In an effort to investigate mechanism of fatty allograft-associated PNFs, we induced fatty liver disease in donor animals by feeding rats a diet deficient in methionine and choline (MCD). We performed OLT with allografts of different grades of hepatic steatosis and compared the results to healthy ones. We assessed liver function by considering serum biochemistries, and investigated genome wide responses following OLT of healthy and fatty allograft-associated PNFs. Furthermore, we performed immunohistochemistry to evaluate markers of oxidative stress and reperfusion injury, inflammation, glycolysis and gluconeogenesis, lactate transport, and its utilization as part of the Cori cycle. Strikingly, PNFs are strictly lipid content dependent. Nonetheless, a fat content of ≤17% and an increase in the size of hepatocytes of ≤11% (ballooning) greatly improved outcome of OLTs and the hepatic microcirculation. Mechanistically, PNFs arise from a dysfunctional Cori cycle with complete ablation of the lactate transporter SLC16A1. Thus, lipid-laden hepatocytes fail to perform gluconeogenesis via lactate reutilization, and the resultant hyperlactatemia and lactic acidosis causes cardiac arrhythmogenicity and death. Furthermore, the genomic and immunohistochemistry investigations underscore a dysfunctional Krebs cycle with impaired energy metabolism in lipid-burdened mitochondria. Together, we show fatty allografts to be highly vulnerable towards ischemia/reperfusion-injury, and stabilizing the Cori cycle is of critical importance to avert PNFs.
原位肝移植(OLT)是一种救命手术。然而,移植物可能会因原发性无功能(PNF)而失败。过去,我们发现 PNF 主要与脂肪供体有关,由于其不可预测的性质,迫切需要开发一种疾病模型。为了研究脂肪供体相关 PNF 的机制,我们通过用缺乏蛋氨酸和胆碱的饮食喂养大鼠来诱导供体动物的脂肪肝疾病(MCD)。我们用不同程度肝脂肪变性的同种异体移植物进行 OLT,并将结果与健康移植物进行比较。我们通过考虑血清生化指标来评估肝功能,并在健康和脂肪供体相关 PNF 的 OLT 后研究全基因组反应。此外,我们进行了免疫组织化学以评估氧化应激和再灌注损伤、炎症、糖酵解和糖异生、乳酸转运及其作为科里循环一部分的利用的标志物。引人注目的是,PNF 严格依赖于脂质含量。尽管如此,脂肪含量≤17%和肝细胞大小增加≤11%(气球样变)极大地改善了 OLT 和肝微循环的结果。从机制上讲,PNF 源自功能失调的科里循环,乳酸转运蛋白 SLC16A1 完全被消融。因此,富含脂肪的肝细胞无法通过乳酸再利用进行糖异生,由此产生的高乳酸血症和乳酸性酸中毒导致心律失常和死亡。此外,基因组学和免疫组织化学研究强调了负担过重的线粒体中功能失调的克雷布斯循环和受损的能量代谢。总之,我们发现脂肪供体非常容易受到缺血/再灌注损伤的影响,稳定科里循环对于避免 PNF 至关重要。