单人直视下改良建立大鼠原位肝移植模型

doi:10.3877/cma.j.issn.2095-3232.2017.02.013

目的

探讨简便、有效的大鼠原位肝移植模型建立方法。

方法

基于"二袖套法"，采用单人直视下改良建立大鼠原位肝移植模型的方法对30对SD大鼠行不重建肝动脉的原位肝移植手术。对麻醉、供肝获取、供肝灌注、修肝、受体肝脏切除、受体胆道重建等重要操作步骤进行改良，无肝期受体采用小剂量肝素化方案。观察并记录肝移植手术过程中供肝灌注前供体手术时间、供肝冷缺血时间、无肝期时间、受体手术时间和肝移植手术总时间。术后对30只受体按随机数字表法随机分为A、B两组，其中A组10只，用于检测肝功能；B组20只，用于观察生存情况。生存分析采用Kaplan-Meier生存曲线。

结果

灌注后供肝良好，受体大鼠移植术后一般状况良好，手术过程中供肝灌注前供体手术时间、供肝冷缺血时间、无肝期时间、受体手术时间、肝移植手术总时间分别为（18.5±1.6）、（75.1±3.8）、（20.5±1.8）、（58.3±3.1）、（118.0±4.2）min。A组大鼠肝移植术后2周肝功能逐渐好转，B组大鼠术后1周生存率为100％，2周为90％，1个月为75％。

结论

单人直视下改良建立大鼠原位肝移植模型是一种简便、有效的方法。

关键词: 肝移植, 大鼠, 模型，动物

Objective

To investigate a convenient and efficient approach for the establishment of rat models with orthotopic liver transplantation (LT).

Methods

Based on the two-cuff technique, orthotopic LT without hepatic artery reconstruction was performed on 30 pairs of Sprague-Dawley (SD) rats through the modified approach for establishment of rat models with orthotopic LT under direct vision of single operator. The key procedures including the anesthesia, donor liver procurement, donor liver perfusion, liver repairing, liver resection of recipient and biliary tract reconstruction of recipient were modified. A small dose of heparinization was administered in the recipients during the anhepatic phase. During the process of LT, the operation time of the donor before donor liver perfusion, cold ischemia time of donor liver, time of anhepatic phase, operation time of the recipient and total time of LT were observed and recorded. After the surgery, 30 recipients were randomly divided into group A (n=10) and group B (n=20) according to the random number table method. The rats in group A were used for detecting the liver function, while rats in group B were used for observing the survival status. Survival analysis was performed using Kaplan-Meier survival curve.

Results

The status of the donor liver was well after liver perfusion. The general status of the recipients was fine after LT. During the process of LT, the operation time of the donor before donor liver perfusion, cold ischemia time of donor liver, time of anhepatic phase, operation time of the recipient and total time of LT was respectively (18.5±1.6), (75.1±3.8), (20.5±1.8), (58.3±3.1) and (118.0±4.2) min. Liver function was gradually getting better 2 weeks after LT in group A. The 1-week, 2-week, 1-month survival rate after surgery was respectively 100%, 90%, and 75% in group B.

Conclusion

The modified establishment of rat models with orthotopic LT under direct vision of single operator is a convenient and efficient approach.

Key words: Liver transplantation, Rats, Models, animal

图3 肝脏切除及植入　注：3a为结扎受体肝动脉；3b为缝合供体和受体肝上下腔静脉；3c为供体和受体门静脉进行套管吻合

表1 肝移植术后A组大鼠各时间点的肝功能指标（±s）

术后时间（d）	鼠数（只）	肝功能指标
术后时间（d）	鼠数（只）	AST（U/L）	ALT（U/L）	TB （μmol/L）
3	10	508±30	237±15	5.8±0.5
7	10	237±15	126±15	4.1±0.6
14	10	137±12	77±9	2.2±0.3

表1 肝移植术后A组大鼠各时间点的肝功能指标（±s）

术后时间（d）	鼠数（只）	肝功能指标
术后时间（d）	鼠数（只）	AST（U/L）	ALT（U/L）	TB （μmol/L）
3	10	508±30	237±15	5.8±0.5
7	10	237±15	126±15	4.1±0.6
14	10	137±12	77±9	2.2±0.3

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Modified approach for establishment of rat models with orthotopic liver transplantation under direct vision of single operator

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Modified approach for establishment of rat models with orthotopic liver transplantation under direct vision of single operator