大鼠心脏死亡器官捐献供体原位肝移植模型建立的经验总结

武睿超; 黄兆宇; 张黎; 刘钧汉; 郑克谱; 冉江华

doi:10.3969/j.issn.1674-7445.2018.04.011

大鼠心脏死亡器官捐献供体原位肝移植模型建立的经验总结

doi: 10.3969/j.issn.1674-7445.2018.04.011

650500 昆明医科大学附属甘美医院暨昆明市第一人民医院

基金项目:

云南省应用基础研究 2015FA010

云南省卫生科技计划项目 2014NS200

详细信息

作者简介:
武睿超，女, 1990年生，硕士研究生，研究方向为肝移植，Email：wrcxg69@163.com

通讯作者:
冉江华，男，1965年生，博士，主任医师，研究方向为肝胆胰外科及肝移植，Email:rjh2u@163.com

中图分类号: R617, R-332
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出版历程
- 收稿日期: 2018-04-28
- 网络出版日期: 2021-01-19
- 刊出日期: 2018-07-15

Summary of experience in the establishment of rat models with orthotopic liver transplantation from donation after cardiac death

The First People' s Hospital of Kunming and Gan Mei Hospital affiliated to Kunming Medical University, Kunming 650500, China

More Information

Corresponding author: Ran Jianghua, Email:rjh2u@163.com

摘要

摘要: 目的探讨建立大鼠心脏死亡器官捐献（DCD）供体原位肝移植模型的技巧并总结经验。方法将120只大鼠按热缺血时间分为3组：A组（热缺血0 min）、B组(热缺血10 min)、C组（热缺血20 min），每组各40对。通过改良“二袖套”法，对3组大鼠行原位肝移植术，记录3组大鼠手术各阶段所用时间。记录3组大鼠手术结束时，术后24 h、72 h、7 d的存活率，如出现死亡及时解剖分析死亡原因。结果 3组大鼠的供肝冷缺血时间、无肝期及受体手术时间的差异均无统计学意义（均为P > 0.05）。手术结束时，A组、B组、C组大鼠的存活率分别为97%、97%、100%；术后24 h，3组相应存活率分别为92%、90%、92%；术后72 h，3组相应存活率分别为90%、80%、77%；术后7 d，3组相应存活率分别为85%、70%、57%。3组大鼠手术结束、术后24 h和术后72 h存活率比较，差异均无统计学意义（均为P > 0.05），术后7 d C组大鼠的存活率低于A组，差异有统计学意义（P < 0.05）。术中和术后24 h内死亡原因多为手术操作导致，术后72 h死亡原因多为胆漏和缺血性肝衰竭，术后7 d死亡原因多为胆道并发症，且随着热缺血时间的延长，胆道并发症的发生只数增多。结论大鼠DCD供体原位肝移植模型的稳定建立关键在于保护肝脏和胆道功能，难点在于肝上下腔静脉的吻合和缩短无肝期。
- 心脏死亡器官捐献 /
- 供体 /
- 原位肝移植 /
- 大鼠模型 /
- 热缺血时间 /
- 无肝期 /
- 胆道并发症 /
- 肝衰竭
Abstract: Objective To explore the skills and summarize the experience in the establishment of orthotopic liver transplantation rat models from donation after cardiac death (DCD). Methods According to the time of warm ischemia, 120 rats were divided into 3 groups: group A (warm ischemia for 0 min, n=40 pairs), group B (warm ischemia for 10 min, n=40 pairs) and group C (warm ischemia for 20 min, n=40 pairs). Orthotopic liver transplantation was performed by the modified two-cuff technique in 3 groups. The time of each stage of surgery was recorded in 3 groups. The survival rate at the end of surgery, 24 h, 72 h and 7 d after surgery was recorded in 3 groups. The dead rats were immediately subject to anatomical examination to identify the cause of death. Results The cold ischemia time of donor liver, anhepatic phase and operation time of the recipients did not significantly differ among three groups (all P > 0.05). In groups A, B and C, the survival rate at the end of surgery was 97%, 97%, and 100% respectively. The survival rate at postoperative 24 h was 92%, 90% and 92% respectively. The survival rate at postoperative 72 h was 90%, 80% and 77% respectively. The survival rate at postoperative 7 d was 85%, 70% and 57% respectively. The survival rate at the end of surgery, postoperative 24 h and 72 h did not significantly differ among 3 groups (all P > 0.05). At postoperative 7 d, the survival rate in group C was significantly lower than that in group A (P < 0.05). Surgical operation was the major cause of intraoperative and postoperative 24 h death. Bile leakage and ischemic hepatic failure were the causes of death at postoperative 72 h. Biliary duct complications were the main causes of death at postoperative 7 d. The quantity of rats developing with biliary duct complications was increased along with the prolongation of warm ischemic time. Conclusions The success of stable establishment of rat models with orthotopic liver transplantation from DCD depends upon the protection of the liver and biliary function. The difficulty lies in the anastomosis of the suprahepatic inferior vena cava and the shortening of anhepatic phase.
- Donation after cardiac death /
- Donor /
- Orthotopic liver transplantation /
- Rat model /
- Warm ischemic time /
- Anhepatic phase /
- Biliary complication /
- Hepatic failure

HTML全文

图 1 大鼠原位肝移植的手术过程

A图为夹闭心脏基底部，开始热缺血时间计时；B图为供肝置入胆管支撑管；C图为供肝完成套管；D图为受体夹闭门静脉、肝上下腔静脉，进入无肝期，经门静脉驱血；E图为受体肝上下腔静脉吊线完成开始缝合；F图为完成肝上下腔静脉吻合；G图为受体门静脉套管吻合；H图为受体胆管支撑管置入吻合；I图为受体门静脉、肝下下腔静脉、胆管吻合完毕，肝脏复血良好

Figure 1. The operative process of orthotopic liver transplantation in rats

下载: 全尺寸图片幻灯片

表 1 3组大鼠手术各阶段的时间

Table 1. The time of each stage of operation of rats in three groups(x±s, min)

组别	n	供肝冷缺血时间	无肝期	受体手术时间
A组	40	48.1±3.7	20.6±2.6	54.3±2.2
B组	40	47.3±3.6	20.7±2.7	54.7±2.2
C组	40	47.7±3.7	20.2±2.5	54.7±2.5

下载: 导出CSV

表 2 3组大鼠术后各时间点存活率比较

Table 2. Comparison of survival rates of rats among three groups at each time point after surgery [n (%)]

组别	n	存活率
组别	n	手术结束	术后24 h	术后72 h	术后7 d
A组	40	39(97)	37(92)	36(90)	34(85)
B组	40	39(97)	36(90)	32(80)	28(70)
C组	40	040(100)	37(92)	31(77)	23(57)a
与A组比较，^aP < 0.05

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