脑心双死亡器官捐献肝损伤状态犬体外膜式氧合模型的建立

doi:10.3877/cma.j.issn.2096-1537.2017.02.008

中华重症医学电子杂志 ›› 2017, Vol. 03 ›› Issue (02) : 116 -121. doi: 10.3877/cma.j.issn.2096-1537.2017.02.008

所属专题：文献；

基础研究

脑心双死亡器官捐献肝损伤状态犬体外膜式氧合模型的建立

朱艳平¹, 曹璐², 姚纪友¹, 唐云华³, 胡晓光¹, 蔡常洁¹, 童荔¹^,^†(

)

^1. 510080　广州，中山大学附属第一医院重症二科
^2. 510080　广州，中山大学附属第一医院体外循环科
^3. 510080　广州，中山大学附属第一医院器官移植科

收稿日期:2017-05-03 出版日期:2017-05-28
通信作者: 童荔
基金资助:
广东省科技计划项目(20130607c)

Extracorporeal membrane oxygenation model establishment of dogs on basis of donation after brain-cardiac death with liver injury

Yanping Zhu¹, Lu Cao², Jiyou Yao¹, Yunhua Tang³, Xiaoguang Hu¹, Changjie Cai¹, Li Tong¹^,^†()

^1. The 2nd Department of Critical Care Medicine, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China
^2. Department of Extracorporeal Circulation, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China
^3. Department of Organ Transplantation, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China

Received:2017-05-03 Published:2017-05-28
Corresponding author: Li Tong
About author:
Corresponding author: Tong Li, Email: 407632428@qq.com

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引用本文：

朱艳平, 曹璐, 姚纪友, 唐云华, 胡晓光, 蔡常洁, 童荔. 脑心双死亡器官捐献肝损伤状态犬体外膜式氧合模型的建立[J]. 中华重症医学电子杂志, 2017, 03(02): 116-121.

Yanping Zhu, Lu Cao, Jiyou Yao, Yunhua Tang, Xiaoguang Hu, Changjie Cai, Li Tong. Extracorporeal membrane oxygenation model establishment of dogs on basis of donation after brain-cardiac death with liver injury[J]. Chinese Journal of Critical Care & Intensive Care Medicine(Electronic Edition), 2017, 03(02): 116-121.

目的

在稳定的犬脑心双死亡器官捐献（DBCD）肝损伤基础上，建立体外膜式氧合（ECMO）循环系统。

方法

20只比格犬，首先采用缓慢颅内加压法诱导脑死亡，确认脑死亡后，放置ECMO动静脉插管，采用Pringle法，阻断门静脉和肝动脉血流60 min，造成DBCD肝损伤模型，连接ECMO建立循环系统。采集阻断前后血标本进行血气分析，检测乳酸、钠离子、钾离子、钙离子、血糖浓度，同时检测血清丙氨酸转氨酶（ALT）、天门冬氨酸转氨酶（AST）、乳酸脱氢酶（LDH）浓度和肿瘤坏死因子α（TNF-α）、白介素6（IL-6）浓度，并采用Student′s t检验比较。

结果

20只犬脑死亡判定均成功。2只犬在建立肝损伤时，开放阻断钳后心跳骤停死亡，余18只均在DBCD肝损伤基础上成功建立ECMO循环系统，成功率为90.0%（18/20）。与阻断前比较，肝脏出现明显的缺血缺氧表现，乳酸浓度上升［（4.57±0.35）mmol/L vs （1.18±0.15）mmol/L］，且差异有统计学意义（t=8.91，P<0.001）；ALT、AST、LDH浓度均上升［（400.40±15.13）U/L vs （27.40±1.98）U/L，（444.75±19.65）U/L vs （25.40±0.98）U/L，（274.00±19.54）U/L vs （55.17±6.26）U/L］，且差异均有统计学意义（t=24.45、21.32、10.67，P均<0.001）；TNF-α、IL-6浓度也均升高［（29.23±1.85）pg/ml vs （8.37±1.30）pg/ml，（5.54 ±1.11）pg/ml vs （2.25±0.81）pg/ml］，且差异有统计学意义（t=9.21，P<0.001；t=2.39，P=0.03）。

结论

本研究成功建立脑心双死亡肝损伤状态下ECMO循环系统动物模型，为DBCD肝损伤ECMO修复研究奠定基础。

关键词: 脑心双死亡, 肝损伤, 体外膜肺氧合, 动物模型

Objective

To establish extracorporeal membrane oxygenation (ECMO) circulatory system on the basis of donation after brain death plus cardiac death (DBCD) with liver injury in dogs.

Methods

Brain death model via increasing intracranial pressure gradually were replicated in 20 male dogs. After confirming the brain death, femoral arteriovenous ECMO catheters were inserted. Pringle-Maneuver was used to block the portal vein and hepatic artery flow for 60 minutes. Extracorporeal membrane oxygenation system was established on the basis of DBCD with liver injury. Levels of lactate, Na⁺, K⁺, Ca²⁺, Glucose, alanine aminotransferase (ALT), aspartic transaminase (AST), lactic dehydrogenase (LDH), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) before and after the blockade was measured. Student′s t test was used to analyze the data.

Results

Brain death model were replicated successfully in all 20 dogs, among which 2 died of cardiac arrest after clamp removed. ECMO system was established successfully in 18 (90.0%) dogs. 60 minutes after blockade, the liver appeared obvious hypoxic-ischemic manifestation. Blood lactate concentration was significant higher after blockade [(4.57±0.35) mmol/L vs (1.18±0.15) mmol/L, t=8.91, P<0.001] as well as the level of ALT, AST and LDH [(400.40±15.13) U/L vs (27.40±1.98) U/L, (444.75±19.65) U/L vs (25.40±0.98) U/L, (274.00±19.54) U/L vs (55.17±6.26) U/L, t=24.45, 21.32, 10.67, all P<0.001]. The TNF-α and IL-6 concentration were also significant higher after blockade compare to the baseline [(29.23±1.85) pg/ml vs (8.37±1.30) pg/ml, t=9.21, P<0.001; (5.54±1.11) pg/ml vs (2.25±0.81) pg/ml, t=2.39, P=0.03].

Conclusion

This study successfully established a liver injury model with brain death, which provides an ideal model to investigate the protection of ECMO on DBCD donors with liver injury.

Key words: Brain-cardiac death, Liver injury, Extracorporeal membrane oxygenation, Animal model

图1，2 脑死亡诱导前后脑电图。图1示诱导前，当疼痛刺激时脑电活动出现强烈波动反应；图2示诱导后静息状态，给予疼痛刺激时脑电图波形未见明显波动

图4 肝脏阻断过程中，肝脏颜色变化为暗红色，肿胀

表1 第一肝门阻断前后犬血气分析指标变化（mmol/L，±s）

时间	只数	乳酸	钠离子	钾离子	钙离子	血糖
阻断前	18	1.18±0.15	145.20±0.66	3.67±0.15	1.17±0.02	6.60±0.29
阻断后	18	4.57±0.35	144.00±1.07	3.75±0.16	1.18±0.01	7.51±0.34
t值		8.91	0.97	0.38	0.19	2.02
P值		<0.001	0.33	0.71	0.85	0.49

表1 第一肝门阻断前后犬血气分析指标变化（mmol/L，±s）

时间	只数	乳酸	钠离子	钾离子	钙离子	血糖
阻断前	18	1.18±0.15	145.20±0.66	3.67±0.15	1.17±0.02	6.60±0.29
阻断后	18	4.57±0.35	144.00±1.07	3.75±0.16	1.18±0.01	7.51±0.34
t值		8.91	0.97	0.38	0.19	2.02
P值		<0.001	0.33	0.71	0.85	0.49

表2 第一肝门阻断前后犬肝功能指标、炎症因子浓度变化（±s）

时间	只数	肝功能指标			炎症因子
时间	只数	ALT（U/L）	AST（U/L）	LDH（U/L）	TNF-α （pg/ml）	IL-6 （pg/ml）
阻断前	18	27.40±1.98	25.40±0.98	55.17±6.26	8.37±1.30	2.25±0.81
阻断后	18	400.40±15.13	444.75±19.65	274.00±19.54	29.23±1.85	5.54±1.11
t值		24.45	21.32	10.67	9.21	2.39
P值		<0.001	<0.001	<0.001	<0.001	0.03

表2 第一肝门阻断前后犬肝功能指标、炎症因子浓度变化（±s）

时间	只数	肝功能指标			炎症因子
时间	只数	ALT（U/L）	AST（U/L）	LDH（U/L）	TNF-α （pg/ml）	IL-6 （pg/ml）
阻断前	18	27.40±1.98	25.40±0.98	55.17±6.26	8.37±1.30	2.25±0.81
阻断后	18	400.40±15.13	444.75±19.65	274.00±19.54	29.23±1.85	5.54±1.11
t值		24.45	21.32	10.67	9.21	2.39
P值		<0.001	<0.001	<0.001	<0.001	0.03

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