Berardo Clarissa, Di Pasqua Laura Giuseppina, Siciliano Veronica, Rizzo Vittoria, Richelmi Plinio, Ferrigno Andrea, Vairetti Mariapia
Department of Internal Medicine and Therapeutics, Unit of Cellular and Molecular Pharmacology and Toxicology, University of Pavia, Pavia, Italy.
Department of Molecular Medicine, Fondazione IRCCS Policlinico S. Matteo and University of Pavia, Pavia, Italy.
Ann Transplant. 2017 Sep 29;22:581-589. doi: 10.12659/aot.904631.
BACKGROUND Ischemic cholangitis is the main cause of liver failure after transplantation and subnormothermic machine perfusion may represent a better strategy than conventional cold storage, minimizing preservation injury. We compared livers preserved by machine perfusion at 20°C (MP20) or by cold storage at 4°C (CS4) with regard to hypoxia-inducible factor (HIF)-1α mRNA expression and protein stabilization in hypoxic conditions. MATERIAL AND METHODS Livers from male Wistar rats were stored on ice at 4°C in UW solution (CS4) or perfused with oxygenated Krebs-Henseleit buffer at 20°C (MP20) for six hours. After preservation, the livers were reperfused for two hours with oxygenated Krebs-Henseleit buffer at 37°C to simulate reimplantation. We collected bile, perfusate, and tissue samples. Transaminases, lactate dehydrogenase, bilirubin, and lactic acid were assayed in the perfusate and bile. ATP/ADP, glycogen, HIF-1α mRNA, and protein expression were measured in the tissue homogenates. RESULTS At the end of preservation, as well as after reperfusion, HIF-1α mRNA expression was significantly higher in the ischemic CS4 livers. Although the hypoxic conditions found in CS4 preservation stabilized HIF-1α protein was significantly higher in the CS4 livers at the end of preservation, no difference was observed after reperfusion, likely because of the oxygen in the reperfusion medium. After reperfusion, the MP20 livers released less transaminases and LDH. The MP20 livers had higher ATP/ADP, glycogen, and biliary bilirubin after both preservation and reperfusion when compared with the CS4 livers. CONCLUSIONS The data demonstrated that MP20 was associated with a lower HIF-1α expression and organ injury with respect to CS4, suggesting that oxygen provided by this preservation setting might approximate the organ request, thus avoiding the ischemic injury usually observed during organ preservation by cold storage.
缺血性胆管炎是移植后肝衰竭的主要原因，亚低温机器灌注可能比传统冷藏是更好的策略，可将保存损伤降至最低。我们比较了在20°C下通过机器灌注（MP20）或在4°C下冷藏（CS4）保存的肝脏在缺氧条件下缺氧诱导因子（HIF）-1α mRNA表达和蛋白质稳定性。材料与方法：雄性Wistar大鼠的肝脏在4°C的UW溶液中冰存（CS4）或在20°C下用含氧的克氏-亨氏缓冲液灌注（MP20）6小时。保存后，肝脏在37°C下用含氧的克氏-亨氏缓冲液再灌注2小时以模拟再植入。我们收集了胆汁、灌注液和组织样本。测定灌注液和胆汁中的转氨酶、乳酸脱氢酶、胆红素和乳酸。在组织匀浆中测量ATP/ADP、糖原、HIF-1α mRNA和蛋白质表达。结果：在保存结束时以及再灌注后，缺血性CS4肝脏中HIF-1α mRNA表达显著更高。尽管在CS4保存中发现的缺氧条件使HIF-1α蛋白在保存结束时在CS4肝脏中显著更高，但再灌注后未观察到差异，可能是因为再灌注介质中的氧气。再灌注后，MP20肝脏释放的转氨酶和LDH较少。与CS4肝脏相比，MP20肝脏在保存和再灌注后均具有更高的ATP/ADP、糖原和胆汁胆红素。结论：数据表明，与CS4相比，MP20与较低的HIF-α表达和器官损伤相关，表明这种保存设置提供的氧气可能接近器官需求，从而避免了通常在冷藏器官保存期间观察到的缺血性损伤。
