间充质干细胞体外修复DCD供肝的研究

A study for mesenchymal stem cell repairing DCD graft during preservation

目的探讨体外心脏死亡器官捐献(DCD)供肝的修复方法,采用大鼠骨髓间充质干细胞(BMMSCs)联合常温机械灌注(NMP),研究其对DCD肝脏的影响。方法体外培养BMMSCs,建立心脏死亡后热缺血45分钟的模型。将Wistar大鼠随机分成3组:A组(冷保存组)、B组(单纯NMP组)和C组(NMP+BMMSCs组),4小时为各组研究时间点(n=5)。血生化仪检测循环液中的丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST),苏木素-伊红(HE)染色观察肝脏的病理情况,血气分析仪检测灌注组的耗氧量。结果 C组ALT和AST的量明显低于B组(ALT:F=177.98,P<0.05;AST:F=284.38,P<0.05);C组ALT和AST的量明显低于A组(ALT:F=3 100.39,P<0.05;AST:F=192.88,P<0.05)。C组的耗氧量在1小时后明显高于B组(4小时:F=22.90,P<0.05)。肝脏病理显示C组的修复作用明显优于其他两组。结论 BMMSCs联合NMP对DCD肝脏功能、病理及细胞活性均具有明显保护作用,为修复DCD供肝提供了新的方法。

Objective To explore the method of repairing donor liver from donation after cardiac death（DCD） in vitro, using BMMSCs combined with normal mechanical perfusion（NMP）at room temperature to study the effect on the liver after cardiac death. Methods BMMSCs were cultured in vitro and the DCD rat model was established with a warm ischemia time of 45 min after cardiac death. Wistar rats were randomly divided into 3 groups： group A（cold storage）, group B（NMP alone） and group C（NMP ＋ BMMSCs）. All the livers were preserved for 4 h（n = 5）. Alanine aminotransferase（ALT） and aspartate transaminase（AST） in the circulating fluid were detected by a blood biochemical instrument, and the histological presentations of the livers were showed by a hematoxylineosin（HE） staining. Oxygen consumption was measured by a blood gas analyzer. Results The levels of ALT and AST in group C were significantly lower than those in group B（ALT ：t = 177.98,P〈0.05;AST ：t = 284.38, P〈0.05） or group A（ALT ：t = 3 100.39,P〈0.05;AST ：t = 192.88,P〈0.05）. The oxygen consumption of group C was significantly higher than that of group B after preservation for 1 h（4 h ：t = 22.90,P〈0.05）. Histological examination of liver showed that the histological presentation of group C was impoved in comparison with the other two groups. Conclusion The combining of BMMSCs and NMP has an obvious protective effect on the DCD liver, which was showed by ALT,AST,histological presentation and cell activity. It may serve as a new method to repair DCD liver graft.