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

[1]	Dutkowski P, Linecker M, DeOliveira ML, et al. Challenges to liver transplantation and strategies to improve outcomes[J]. Gastroenterology, 2015, 148(2):307-323.
[2]	Jadlowiec CC, Taner T. Liver transplantation: current status and challenges[J]. World J Gastroenterol, 2016, 22(18):4438-4445.
[3]	Zhai Y, Petrowsky H, Hong JC, et al. Ischaemia-reperfusion injury in liver transplantation--from bench to bedside[J]. Nat Rev Gastroenterol Hepatol, 2013, 10(2):79-89.
[4]	Kashfi A, Mehrabi A, Pahlavan PS, et al. A review of various techniques of orthotopic liver transplantation in the rats[J]. Transplant Proc, 2005, 37(1):185-188.
[5]	尹东亮，孙翀，朱焕斌，等．CTLA4-Ig基因修饰骨髓间充质干细胞抑制大鼠肝移植排斥反应[J]．器官移植，2014，5(4),231-236.
[6]	Qin L, Guan HG, Zhou XJ, et al. Blockade of 4-1BB/4-1BB ligand interactions prevents acute rejection in rat liver transplantation[J]. Chin Med J, 2010, 123(2):212-215.
[7]	Fujiki M, Esquivel CO, Martinez OM, et al. Induced tolerance to rat liver allografts involves the apoptosis of intragraft T cells and the generation of CD4(+)CD25(+)FoxP3(+) T regulatory cells[J]. Liver Transpl, 2010, 16(2):147-154.
[8]	Lee S, Chatters AC, Chandler JG, et al. A technique for orthotopic liver transplantation in the rat[J]. Transplantation, 1973, 16(6):664-669.
[9]	Kamada N, Calne RY. Orthotopic liver transplantation in the rat. technique using cuff for portal vein anastomosis and biliary drainage[J]. Transplantation, 1979, 28(1):47-50.
[10]	沈浩元，王炜，刘猛，等．改良单人操作大鼠原位肝移植模型的建立[J]．中国普外基础与临床杂，2011，18(8):825-828.
[11]	Oldani G, Lacotte S, Morel P, et al. Orthotopic liver transplantation in rats[J]. J Vis Exp, 2012(65):4143.
[12]	王浩，温晓薇，陈欢直，等．改良套管法大鼠肝移植模型的建立[J]．肿瘤研究与临床，2016，28(4):262-265.
[13]	Aller MA, Arias N, Prieto I, et al. A half century (1961-2011) of applying microsurgery to experimental liver research[J]. World J Hepatol, 2012, 4(7):199-208.
[14]	Vdoviaková K, Petrovová E, Maloveská M, et al. Surgical anatomy of the gastrointestinal tract and its vasculature in the laboratory rat[J]. Gastroenterol Res Pract, 2016:2632368.
[15]	Martins PN, Neuhaus P. Surgical anatomy of the liver, hepatic vasculature and bile ducts in the rat[J]. Liver Int, 2007, 27(3):384-392.
[16]	史冀华，朱盛兴，张水军．大鼠肝部分切除术的应用解剖及实施[J]．世界华人消化杂志，2008，16(22):2516-2520.
[17]	吕毅，潘承恩，孟令忠，等．SD大鼠肝脏及附属管道的应用解剖[J]．西安医科大学学报，1998，19(1):70-73.
[18]	罗刚健，赵伟成，朱国松，等．大鼠自体肝移植模型的改良——附216例分析[J]．实用医学杂志，2012，28(10):1605-1607.
[19]	许勇刚，翁明哲，张金彦，等．单人建立大鼠原位肝移植模型的手术体会[J]．现代生物医学进展，2012，12(16):3026-3028,3044.
[20]	时军，吴勤荣，罗文峰，等．大鼠原位肝移植模型的建立[J]．中国组织工程研究，2012，16(5):761-765．
[21]	Schemmer P, Schoonhoven R, Swenberg JA, et al. Gentle in situ liver manipulation during organ harvest decreases survival after rat liver transplantation: role of Kupffer cells[J]. Transplantation, 1998, 65(8):1015-1020.
[22]	Tokunaga Y, Ozaki N, Wakashiro S, et al. Effects of perfusion pressure during flushing on the viability of the procured liver using noninvasive fluorometry[J]. Transplantation, 1988, 45(6):1031-1035.

[1]	施洪鑫, 唐志方, 郭民政, 浦路桥, 齐保闯, 任俊筱, 李川. 碘乙酸钠在鼠类骨关节炎造模的方法和作用探索[J]. 中华关节外科杂志(电子版), 2023, 17(04): 534-539.
[2]	张晓燕, 肖东琼, 高沪, 陈琳, 唐发娟, 李熙鸿. 转录因子12过表达对脓毒症相关性脑病大鼠大脑皮质的保护作用及其机制[J]. 中华妇幼临床医学杂志(电子版), 2023, 19(05): 540-549.
[3]	李坤河, 寇萌佳, 邝立挺. 肝移植术后二次气管插管的危险因素及预测模型的建立[J]. 中华普通外科学文献(电子版), 2023, 17(05): 366-371.
[4]	中华医学会器官移植学分会, 中国医师协会器官移植医师分会, 上海医药行业协会. 中国肝、肾移植受者霉酚酸类药物应用专家共识(2023版)[J]. 中华移植杂志(电子版), 2023, 17(05): 257-272.
[5]	陆闻青, 陈昕怡, 任雪飞. 遗传代谢病儿童肝移植受者术后生活质量调查研究[J]. 中华移植杂志(电子版), 2023, 17(05): 287-292.
[6]	范铁艳, 李君, 陈虹. 肝移植术后新发戊型病毒性肝炎的诊治经验[J]. 中华移植杂志(电子版), 2023, 17(05): 293-296.
[7]	陈朔, 陈峰, 程飞, 项捷. 糖原累积病Ⅰ型并发胰腺炎肝移植术后胰腺梗死一例[J]. 中华移植杂志(电子版), 2023, 17(05): 300-302.
[8]	汤鹏昊, 张武. 肠道微生态与肝移植围手术期并发症相关研究进展[J]. 中华移植杂志(电子版), 2023, 17(05): 303-307.
[9]	严庆, 刘颖, 邓斐文, 陈焕伟. 微血管侵犯对肝癌肝移植患者生存预后的影响[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 624-629.
[10]	廖梅, 张红君, 金洁玚, 吕艳, 任杰. 床旁超声造影对肝移植术后早期肝动脉血栓的诊断价值[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 630-634.
[11]	李秉林, 吕少诚, 潘飞, 姜涛, 樊华, 寇建涛, 贺强, 郎韧. 供肝灌注液病原菌与肝移植术后早期感染的相关性分析[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 656-660.
[12]	吕垒, 冯啸, 何凯明, 曾凯宁, 杨卿, 吕海金, 易慧敏, 易述红, 杨扬, 傅斌生. 改良金氏评分在儿童肝豆状核变性急性肝衰竭肝移植手术时机评估中价值并文献复习[J]. 中华肝脏外科手术学电子杂志, 2023, 12(06): 661-668.
[13]	王孟龙. 肿瘤生物学特征在肝癌肝移植治疗中的意义[J]. 中华肝脏外科手术学电子杂志, 2023, 12(05): 490-494.
[14]	萨仁高娃, 张英霞, 邓伟, 闫诺, 樊宁. 超声引导下鼠肝消融术后组织病理特征的变化规律及影响[J]. 中华消化病与影像杂志(电子版), 2023, 13(06): 394-398.
[15]	王小红, 钱晶, 翁文俊, 周国雄, 朱顺星, 祁小鸣, 刘春, 王萍, 沈伟, 程睿智, 秦璟灏. 巯基丙酮酸硫基转移酶调控核因子κB信号介导自噬对重症急性胰腺炎大鼠的影响及机制[J]. 中华消化病与影像杂志(电子版), 2023, 13(06): 422-426.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

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

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

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