D-氨基半乳糖诱导的非人灵长类动物急性肝衰竭模型的建立

黄一鸣; 王励; 李进军; 张毅; 陈文捷; 杨扬

doi:10.3969/j.issn.1674-7445.2019.01.007

D-氨基半乳糖诱导的非人灵长类动物急性肝衰竭模型的建立

doi: 10.3969/j.issn.1674-7445.2019.01.007

510630 广州，中山大学附属第三医院肝脏外科暨肝移植中心广东省肝脏疾病研究重点实验室

基金项目:

国家重点研发计划 2017YFA0104304

国家自然科学基金 81570593

国家自然科学基金 81770648

广东省自然科学基金研究团队项目 2015A030312013

广东省科技计划项目 2017A020215023

详细信息

作者简介:
黄一鸣，男，1992年生，硕士研究生，研究方向为肝移植，Email：515003256@qq.com

通讯作者:
杨扬，男，1971年生，教授，主任医师，博士研究生导师，研究方向为肝移植，Email：yysysu@163.com

中图分类号: R617, R575.3
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- 被引次数: 0
出版历程
- 收稿日期: 2018-10-28
- 网络出版日期: 2021-01-19
- 刊出日期: 2019-01-15

Establishment of acute liver failure models induced by D-galactosamine in non-human primates

Department of Hepatic Surgery, Liver Transplantation Center, the Third Affiliated Hospital of Sun Yat-sen University, Guangdong Provincial Key Laboratory of Liver Disease Research, Guangzhou 510630, China

More Information

Corresponding author: Yang Yang, E-mail:yysysu@163.com

摘要

摘要: 目的探讨不同剂量D-氨基半乳糖（D-Gal）对非人灵长类动物食蟹猴的影响并建立不同程度急性肝衰竭（ALF）猴模型。方法将12只食蟹猴分为3组，每组4只，分别为低剂量组、中剂量组和高剂量组，给药剂量分别为0.23、0.25、0.27 g/kg。各组动物均在清醒状态（无麻醉）下，将相应剂量的D-Gal溶液经前臂静脉一次性注射入猴体内。记录猴的存活时间，观察是否出现消化道症状和肝性脑病症状。分别于给药前0 h，给药后12、24、36、48、60、72、96、120、144 h测量生命体征，留取血标本检测丙氨酸转氨酶（ALT）、总胆红素（TB）、凝血酶原时间（PT）、血氨等指标。取肝脏组织行苏木素-伊红（HE）染色观察病理学改变。结果低剂量组猴全部存活，仅出现一过性肝损伤，无肝性脑病症状，给药后60 h，肝功能和凝血指标达峰值，后逐渐恢复，120 h后基本恢复正常；中剂量组猴病程进展稍缓，在出现严重肝功能损伤及肝性脑病症状后逐渐恢复，仅1只死亡；高剂量组猴均出现肝性脑病症状后死亡，存活时间（72±13）h，肝功能严重受损。肝组织病理学检查示，低剂量组肝组织散在肝细胞坏死，炎症细胞浸润；中、高剂量组可见肝小叶结构不清，肝细胞片状坏死伴有明显出血。结论中、高剂量组均符合ALF模型标准，中剂量组ALF程度较轻，有利于肝移植手术等实验的开展；高剂量组ALF程度较重，适用于相关治疗方案疗效评价。
- 非人灵长类动物 /
- 食蟹猴 /
- 急性肝衰竭 /
- 肝性脑病 /
- 肝移植 /
- 肝功能 /
- 凝血 /
- 动物模型
Abstract: Objective To evaluate the effect of different doses of D-galactosamine (D-Gal) on non-human primate cynomolgus monkey and to establish a monkey model with different degree of acute liver failure (ALF). Methods Twelve cynomolgus monkeys were evenly divided into the low-, medium- and high-dose groups (n=4) with a dosage of 0.23, 0.25 and 0.27 g/kg, respectively. In each group, the corresponding dose of D-Gal solution was injected into the monkeys through the forearm vein at one time in a sober state (without anesthesia). The survival time of the cynomolgus monkeys was recorded. Digestive tract and hepatic encephalopathy symptoms were observed. Vital signs were measured at 0 h before and 12, 24, 36, 48, 60, 72, 96, 120 and 144 h after D-Gal administration. Alanine transaminase (ALT), total bilirubin (TB), prothrombin time (PT), blood ammonia and other parameters were detected from the blood samples. The liver tissues were prepared for hematoxylin-eosin (HE) staining to observe the pathological changes. Results All cynomolgus monkeys in the low-dose group survived and transient liver injury was noted without the hepatic encephalopathy symptoms. At 60 h after D-Gal administration, the liver function and coagulation indexes reached the peak, gradually recovered and then basically returned to the normal range at 120 h. In the medium-dose group, the course of disease was relatively slow and gradually recovered after the appearance of severe liver damage and hepatic encephalopathy symptoms and only one animal died. All cynomolgus monkeys in the high-dose group died after developing hepatic encephalopathy symptoms and severe liver damage with a mean survival time of (72±13) h. Pathological examination of liver tissue demonstrated that scattered liver cell necrosis and inflammatory cell infiltration were observed in the liver tissues of the low-dose group. In the medium- and high-dose groups, the hepatic lobule structure was not clear, and the liver cell necrosis in flakes accompanied by evident hemorrhage were documented. Conclusions The D-Gal dosage in the medium- and high-dose groups meet the standards of the ALF model. The degree of ALF in the medium-dose group is relatively slight, which is beneficial to the implementation of liver transplantation. ALF in the high-dose group is relatively severe, which is suitable for the evaluation of the clinical efficacy of therapeutic options.
- Non-human primate animal /
- Cynomolgus monkey /
- Acute liver failure /
- Hepatic encephalopathy /
- Liver transplantation /
- Liver function /
- Coagulation /
- Animal model

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图 1 不同剂量组血清指标的比较

A~D图分别为3组ALT、TB、血氨、PT随时间变化的趋势。与低剂量组比较，^aP < 0.05；与中剂量组比较，^bP < 0.05

Figure 1. Comparison of serum indexes among different dose groups

下载: 全尺寸图片幻灯片

图 2 不同剂量组猴肝组织的病理学图片（HE, ×400）

A~D图分别为正常肝组织、低剂量组、中剂量组、高剂量组的肝组织病理学图片

Figure 2. Histopathological pictures of liver tissue of monkeys in different dose groups

下载: 全尺寸图片幻灯片

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