Tianjin First Central Hospital Clinic Institute, Tianjin Medical University, Tianjin, 300070, People's Republic of China.
Department of Organ Transplantation, Tianjin First Central Hospital, No. 24 Fukang Road, Nankai District, Tianjin, 300192, People's Republic of China.
Cell Tissue Res. 2020 Aug;381(2):239-254. doi: 10.1007/s00441-020-03202-z. Epub 2020 Apr 29.
Donation after circulatory death (DCD) can expand the donor pool effectively. A gap remains in outcome between DCD livers and living donor livers, warranting improved DCD liver quality and urgent resolution. Bone marrow mesenchymal stem cells (BMMSCs) can regulate immunity, participate in the anti-inflammatory response, and secrete cytokines. We investigated the effect of BMMSCs combined with normothermic machine perfusion (NMP) on DCD liver quality, and the role of microcirculation therein. Rat thoracic aortas were clipped to obtain DCD livers, and a rat NMP system was established. The DCD livers were grouped by preservation method: normal, static cold storage (SCS), NMP (P), and BMMSCs plus NMP (BP); storage time was up to 8 h. Liver function in outflow perfusate was detected by biochemical methods; liver tissue histopathology was observed by hematoxylin-eosin staining; hepatocyte ultrastructure was observed by transmission electron microscopy; hepatocyte apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling; liver microcirculation-related indicators were detected by immunofluorescence, immunohistochemistry, Western blotting, and enzyme-linked immunosorbent assay. Compared with SCS, P and BP significantly improved liver function and liver histological damage, reduced hepatocyte apoptosis, and repaired hepatocyte mitochondrial damage after 6 h in vitro. BP also significantly inhibited intrahepatic macrophage activation and intercellular adhesion, improved endothelial damage, and significantly improved endothelin 1-nitric oxide balance and microcirculation perfusion. In conclusion, BP can improve DCD liver microcirculation and quality. The mechanism may be the improvement of improve hepatic sinusoidal endothelial injury and microcirculation perfusion by inhibiting macrophage activation and intercellular adhesion.
在循环死亡（DCD）后捐赠可以有效地扩大供体库。DCD 肝脏和活体供体肝脏之间的结果仍存在差距，需要提高 DCD 肝脏质量并紧急解决。骨髓间充质干细胞（BMMSCs）可以调节免疫，参与抗炎反应并分泌细胞因子。我们研究了 BMMSCs 与常温机器灌注（NMP）联合使用对 DCD 肝脏质量的影响，以及其中微循环的作用。夹闭大鼠胸主动脉以获得 DCD 肝脏，并建立大鼠 NMP 系统。根据保存方法对 DCD 肝脏进行分组：正常、静态冷保存（SCS）、NMP（P）和 BMMSCs 加 NMP（BP）；储存时间长达 8 小时。通过生化方法检测流出灌注液中的肝功能；通过苏木精-伊红染色观察肝组织病理学；通过透射电子显微镜观察肝细胞超微结构；通过末端脱氧核苷酸转移酶 dUTP 缺口末端标记法检测肝细胞凋亡；通过免疫荧光、免疫组织化学、Western blot 和酶联免疫吸附试验检测肝微循环相关指标。与 SCS 相比，P 和 BP 显著改善了肝功能和肝组织损伤，减少了肝细胞凋亡，并在体外 6 小时修复了肝细胞线粒体损伤。BP 还显著抑制了肝内巨噬细胞的激活和细胞间黏附，改善了内皮损伤，并显著改善了内皮素 1-一氧化氮平衡和微循环灌注。总之，BP 可以改善 DCD 肝脏的微循环和质量。其机制可能是通过抑制巨噬细胞激活和细胞间黏附来改善肝窦内皮损伤和微循环灌注。
