Tianjin First Central Hospital Clinic Institute, Tianjin Medical University, Tianjin, People's Republic of China.
Department of Organ Transplantation, Tianjin First Central Hospital, Tianjin, People's Republic of China.
Stem Cells Dev. 2020 Jul 1;29(13):835-852. doi: 10.1089/scd.2019.0301. Epub 2020 May 5.
There is a need to improve the quality of donor liver from donation after circulatory death (DCD). The purpose of this study was to investigate the effects and mechanism of normothermic machine perfusion (NMP) combined with bone marrow mesenchymal stem cells (BMMSCs) on the oxidative stress and mitochondrial function in DCD livers. DCD livers were obtained, a rat NMP system was established, and BMMSCs were extracted and identified. The DCD livers were grouped by their preservation method: Normal, static cold storage (SCS), NMP (P), and NMP combined with BMMSCs (PB), and the preservation time was up to 8 h. An IAR20 cell oxidative stress injury model was established in vitro by simulating DCD oxidative stress injury and coculturing with BMMSCs for 6 h. Compared with SCS group, after 6 h in vitro, the PB and P groups had significantly improved liver function and liver histological damage, reduced hepatocyte apoptosis and oxidative stress, improved hepatocyte mitochondrial damage, and increased mitochondrial membrane potential. These indicators were significantly better in the PB group than in the P group. BMMSCs significantly inhibited reactive oxygen species release from the IAR20 cell oxidative stress model in vitro, ameliorated mitochondrial damage, and increased mitochondrial membrane potential level. BMMSCs also downregulated the JUN N-terminal kinase-nuclear factor kappa B (JNK-NF-κB) signaling pathway significantly in the IAR20 cell oxidative stress model and promoted AMP-activated protein kinase (AMPK) activation. We verified that NMP combined with BMMSCs also played the same role in the PB group. NMP combined with BMMSCs could improve liver quality by relieving oxidative stress injury and improving mitochondrial function in rat DCD livers. The mechanism of protective role might involve inhibiting the JNK-NF-κB pathway to reduce oxidative stress and promote AMPK activation, thereby reducing mitochondrial damage and increase mitochondrial function.
需要提高心脏死亡器官捐献（DCD）供肝的质量。本研究旨在探讨常温机械灌注（NMP）联合骨髓间充质干细胞（BMMSCs）对DCD 肝脏氧化应激和线粒体功能的影响及机制。获取 DCD 供肝，建立大鼠 NMP 系统，提取并鉴定 BMMSCs。根据保存方法将 DCD 供肝分组：正常、静态冷保存（SCS）、NMP（P）和 NMP 联合 BMMSCs（PB），保存时间长达 8 h。体外模拟 DCD 氧化应激损伤，与 BMMSCs 共培养 6 h，建立 IAR20 细胞氧化应激损伤模型。与 SCS 组相比，体外 6 h 后，PB 和 P 组肝功能和肝组织损伤明显改善，肝细胞凋亡和氧化应激减少，肝细胞线粒体损伤改善，线粒体膜电位增加。PB 组各指标明显优于 P 组。BMMSCs 显著抑制 IAR20 细胞氧化应激模型中活性氧的释放，改善线粒体损伤，增加线粒体膜电位水平。BMMSCs 还显著下调 IAR20 细胞氧化应激模型中 JUN 氨基末端激酶-核因子 kappa B（JNK-NF-κB）信号通路，促进 AMP 激活的蛋白激酶（AMPK）激活。我们验证了 NMP 联合 BMMSCs 在 PB 组也发挥了相同的作用。NMP 联合 BMMSCs 可通过减轻氧化应激损伤和改善大鼠 DCD 肝脏线粒体功能来提高肝脏质量。其保护作用的机制可能涉及抑制 JNK-NF-κB 通路减少氧化应激，促进 AMPK 激活，从而减少线粒体损伤和增加线粒体功能。
