小鼠肝动脉重建对长时间冷保存移植肝存活率的影响

目的研究小鼠肝动脉重建(hepatic arterial reconstruction,HAR)对长时间冷保存移植肝存活率的影响。方法同系雄性C57BL/6小鼠68只,分为冷保存时间(cold preservation time,CPT)1 h组、CPT4 h组、CPT8 h组、CPT16 h组、CPT16 h+HAR组(供、受体小鼠各一半)。小鼠供肝经门静脉灌注4℃UW液后保存。肝脏移植采用缝合法(肝上下腔静脉)和袖套法(门静脉、肝下下腔静脉)吻合,胆管采用内支架管重建法。小鼠HAR采用含供体肝动脉的腹主动脉与受体腹主动脉端侧吻合的方法。观察术后5组受体小鼠移植肝的存活时间,用组织学检查肝细胞损伤情况,用免疫组织化学法观察肝细胞再生功能。结果 CPT1 h、4 h、8 h组受体术后12 d移植物的存活率分别为7/7、10/10、9/9。CPT16 h组除1例小鼠存活外,其余均在移植术后36 h内死亡,存活率为10%(1/10),CPT16 h+HAR组受体90%(9/10)存活,两组存活率相比,差异有统计学意义(P0.01)。CPT1 h、4 h组的移植物组织损伤程度轻,CPT8 h组的移植物组织损伤程度较前两组严重,但肝细胞再生活跃。CRT16 h组的移植肝组织表现为广泛肝细胞空泡变性、坏死,肝细胞再生不明显。CPT16 h+HAR组仅有轻度的肝窦淤血,肝细胞空泡变性、坏死改变,肝细胞再生活跃。结论 HAR可提高长时间冷保存移植肝的存活率。

Effect of hepatic arterial reconstruction in mice on the survival rate of liver grafts following long-term cold preservation injury

Objective To study the effect of hepatic arterial reconstruction （HAR） on the survival rate of liver grafts following long-term cold preservation injury. Methods Syngeneie male C57BL/6 mice （ n = 68） were divided into 5 groups： CFF 1, 4, 8, 16 h and CPT 16 h ＋ HAR group. Liver perfusion was flushed into portal vein with 4℃ UW solution and the grafts were preserved in 4~C UW solution. Orthotopic liver transplantation （OLT） was performed with a combination of suture （suprahepatic vena cava） and cuff anastomosis technique （infrahepatic vena cava and portal vein）. The bile duct was reconnected through the intraluminal stent. The methods for HAR in mice include end-to-side anastomosis of abdominal aorta with hepatic artery from donor livers to abdominal aorta in the recipient. Recipient survival was followed up and hepatocellular injury was evaluated by histology. Hepatocyte replication was examined by immunohistochemistry. Results Survival rate on postoperative day 12 in CPT 1, 4, 8 h group was 7/7, 10/10, 9/9, respectively. Recipients （9 of 10） in CPT 16 h group died within 36 h after transplantation, whereas only 10% （1/10） of mice remained alive on day 12 after transplantation. All animals but one in CPT 16 h ＋ HAR group survived 12 d after arterial reconstruction. There was significant difference in survival rate between the two groups （ P 〈 0. 01 ）. Histology showed mild injury in grafts of CPT 1 and 4 h groups. More severe injury and obvious hepatoeyte replication was observed in CPT 8 h group compared with the CPT 1 and 4 h groups. Histology examination in 16 h CPT group showed extensive hepatocyte vacuolar degeneration and areas of necrosis without increased hepatocyte replica- tion. Liver grafts in CPT 16 h ＋ HAR group showed mild sinusoidal congestion, vacuolar degeneration, hepatocyte necrosis and active hepa, ocyte replication. Conclusion HAR can improve the survival rate of liver grafts with severe long-term cold preservation injury.