Department of Surgery, Campus Charité Mitte | Campus Virchow Klinikum, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt- Universität zu Berlin and Berlin Institute of Health, Berlin, Germany.
Institute of Pathology, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt- Universität zu Berlin and Berlin Institute of Health, Berlin, Germany.
PLoS One. 2020 Jul 2;15(7):e0235635. doi: 10.1371/journal.pone.0235635. eCollection 2020.
Normothermic ex vivo liver perfusion (NEVLP) is a promising strategy to increase the donor pool in liver transplantation. Small animal models are essential to further investigate questions regarding organ preservation and reconditioning by NEVLP. A dual vessel small animal NEVLP (dNEVLP) model was developed using metamizole as a vasodilator and compared to conventional portovenous single vessel NEVLP (sNEVLP).
Livers of male Wistar rats were perfused with erythrocyte-supplemented culture medium for six hours by either dNEVLP via hepatic artery and portal vein or portovenous sNEVLP. dNEVLP was performed either with or without metamizole treatment. Perfusion pressure and flow rates were constantly monitored. Transaminase levels were determined in the perfusate at the start and after three and six hours of perfusion. Bile secretion was monitored and bile LDH and GGT levels were measured hourly. Histopathological analysis was performed using liver and bile duct tissue samples after perfusion.
Hepatic artery pressure was significantly lower in dNEVLP with metamizole administration. Compared to sNEVLP, dNEVLP with metamizole treatment showed higher bile production, lower levels of transaminases during and after perfusion as well as significantly lower necrosis in liver and bile duct tissue. Biochemical markers of bile duct injury showed the same trend.
Our miniaturized dNEVLP system enables normothermic dual vessel rat liver perfusion. The administration of metamizole effectively ameliorates arterial vasospasm allowing for six hours of dNEVLP, with superior outcome compared to sNEVLP.
常温体外肝脏灌注（NEVLP）是增加肝移植供体库的一种有前途的策略。小动物模型对于进一步研究通过 NEVLP 进行器官保存和再调理的问题至关重要。本研究开发了一种双血管小动物 NEVLP（dNEVLP）模型，使用美他唑仑作为血管扩张剂，并与传统的门静脉单血管 NEVLP（sNEVLP）进行了比较。
使用含红细胞的培养基通过肝动脉和门静脉进行 dNEVLP 或门静脉 sNEVLP，对雄性 Wistar 大鼠的肝脏进行六小时灌注。dNEVLP 可进行或不进行美他唑仑处理。持续监测灌注压和流量。在灌注开始时以及灌注 3 小时和 6 小时后，测定灌流液中的转氨酶水平。监测胆汁分泌，并每小时测量胆汁 LDH 和 GGT 水平。灌注后，使用肝和胆管组织样本进行组织学分析。
美他唑仑给药的 dNEVLP 肝动脉压显著降低。与 sNEVLP 相比，美他唑仑处理的 dNEVLP 显示出更高的胆汁产量、灌注过程中和灌注结束时更低的转氨酶水平，以及肝和胆管组织的坏死明显减少。胆管损伤的生化标志物也呈现出相同的趋势。
我们的小型化 dNEVLP 系统能够实现常温双血管大鼠肝脏灌注。美他唑仑的给药有效地缓解了动脉血管痉挛，从而实现了 6 小时的 dNEVLP，与 sNEVLP 相比，效果更好。
