Li Tao, Chang Pengkang, Wang Yimeng, Song Yihong, Qu Pengxiang, Wang Bo, Lyu Yi, Hu Liangshuo
Department of Hepatobiliary Surgery, Institute of Advanced Surgical Technology and Engineering, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Department of Urology, Urologic Surgery Center, Xinqiao Hospital, Third Military Medical University (Army Medical University), Chongqing, China.
Sci Rep. 2025 Apr 6;15(1):11762. doi: 10.1038/s41598-025-96265-3.
Living liver transplantation has become a significant and evolving aspect of organ transplantation, with a notable proportion of cases involving pediatric patients. Metabolic-associated fatty liver disease (MAFLD) is the most prevalent chronic liver disease. The growing number of individuals with MAFLD has led to an annual increase in the proportion of non-alcoholic fatty liver donors for pediatric living liver transplantation. Hypothermic oxygenated perfusion (HOPE) has been demonstrated to improve graft quality through the implementation of a continuous mechanical perfusion cycle. However, there is currently a paucity of evidence regarding its ability to reduce steatosis and improve prognosis within a shorter time window of living-organ transplantation, especially in primate models. This study simulated steatotic liver grafts in living liver transplantation using the MAFLD model of the cynomolgus monkey and explored the effects of HOPE combined with the AMPK activator AICAR on the amelioration of the donor liver. The left outer lobe livers were statically cold preserved for two hours, subjected to HOPE for two hours, or treated with HOPE + AICAR (1 mmol/L) for two hours, respectively. Subsequently, a normothermic ex vivo reperfusion model (IRM) simulating post-transplant reperfusion was established using diluted autologous blood. Following simulated reperfusion in vitro, steatotic liver grafts in the static cold preservation group exhibited notable reperfusion injury. The degree of reperfusion injury induced by the remaining two groups was reduced, with the HOPE + AICAR group showing the most significant reduction (P < 0.05). The adenosine triphosphate (ATP) level of the hepatic tissues in the HOPE + AICAR group was observed to improve at two hours of reperfusion, exhibiting a significantly higher level than that in the cold-preserved group (P < 0.05). Furthermore, the HOPE + AICAR group exhibited a notable decline in MDA levels (P < 0.05), accompanied by a considerable reduction in 8-OHdG and lactate concentrations in both the liver tissue and perfusate. Additionally, there was a marked decrease in the release of TNF-α and IL-6 cytokines, along with a reduction in TLR-4 activation (P < 0.05). In comparison to the cold-preserved and HOPE groups, the HOPE + AICAR group demonstrated the capacity to alter the degree of steatosis following a two-hour treatment period, as evidenced by a notable reduction in liver tissue triglyceride and cholesterol levels (P < 0.05). Additionally, p-AMPK levels in liver tissue were significantly increased in the HOPE + AICAR group (P < 0.05). The combination of HOPE and AMPK activators has been shown to reduce the degree of steatotic liver grafts in a relatively short time, significantly reduce reperfusion injury, and improve liver function. This study contributes to the existing body of knowledge on mechanical perfusion in primate models, addressing a previously identified gap in the literature.
活体肝移植已成为器官移植中一个重要且不断发展的领域,其中相当一部分病例涉及儿科患者。代谢相关脂肪性肝病(MAFLD)是最常见的慢性肝病。MAFLD患者数量的增加导致儿科活体肝移植中非酒精性脂肪肝供体的比例逐年上升。低温氧合灌注(HOPE)已被证明可通过实施连续机械灌注循环来提高移植物质量。然而,目前关于其在活体器官移植较短时间窗内减少脂肪变性并改善预后能力的证据尚少,尤其是在灵长类动物模型中。本研究使用食蟹猴的MAFLD模型模拟活体肝移植中的脂肪变性肝移植物,并探讨HOPE联合AMPK激活剂AICAR对供体肝脏改善的影响。左外叶肝脏分别进行两小时的静态冷保存、两小时的HOPE处理或两小时的HOPE + AICAR(1 mmol/L)处理。随后,使用稀释的自体血建立模拟移植后再灌注的常温体外再灌注模型(IRM)。体外模拟再灌注后,静态冷保存组的脂肪变性肝移植物表现出明显的再灌注损伤。其余两组诱导的再灌注损伤程度降低,其中HOPE + AICAR组降低最为显著(P < 0.05)。观察到HOPE + AICAR组肝组织的三磷酸腺苷(ATP)水平在再灌注两小时时有所改善,显著高于冷保存组(P < 0.05)。此外,HOPE + AICAR组的丙二醛(MDA)水平显著下降(P < 0.05),同时肝组织和灌注液中的8-羟基脱氧鸟苷(8-OHdG)和乳酸浓度大幅降低。此外,肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)细胞因子的释放显著减少,同时Toll样受体4(TLR-4)激活降低(P < 0.05)。与冷保存组和HOPE组相比,HOPE + AICAR组在两小时治疗期后显示出改变脂肪变性程度的能力,肝组织甘油三酯和胆固醇水平显著降低(P < 0.05)证明了这一点。此外,HOPE + AICAR组肝组织中的磷酸化AMPK(p-AMPK)水平显著升高(P < 0.05)。已证明HOPE与AMPK激活剂的联合使用可在相对较短时间内降低脂肪变性肝移植物的程度,显著减少再灌注损伤并改善肝功能。本研究为灵长类动物模型中机械灌注的现有知识体系做出了贡献,填补了文献中先前发现的空白。