摘要
目的建立一种操作简单、易开展、存活率高的大鼠急性肝损伤再生修复模型。方法将30只斯泼累格·多雷(Sprague-Dawley,SD)大鼠,按照随机抽样原则将其分为对照组(n=6)和实验组(4个亚组,每组6只,n=24),各组均初次行肝左外叶70%切除手术以模拟急性肝损伤;其中对照组立即于尾缘静脉取血进行生化检测以明确正常肝组织丙氨酸转氨酶(alanine aminotransferase,ALT)、天冬氨酸转氨酶(aspartate aminotransferase,AST)水平,并且立即再次手术切除残存肝左外叶行苏木精-伊红染色(hematoxylin-eosin staining,HE),以明确正常肝组织结构轮廓;各实验亚组分别间隔6 h、12 h、3 d、7 d后于尾缘静脉取血进行ALT、AST生化检测,并且再次手术切除残存肝左外叶进行HE染色,观察各实验亚组大鼠二次术后存活率及状态,以病理学判定标准及两次术后均能长期稳定存活为造模成功的标志。结果 (1)血液生化检测:与对照组相比,实验组于6 h、12 h血清ALT、AST含量显著增高,差异具有统计学意义(P<0.05),且在6 h显著升高[(200.150±1.2357)U/L,(257.483±1.1940)U/L],12 h继续升高[(449.933±1.4949)U/L,(755.700±1.0040)U/L],第3天开始降低[(55.87±0.88)U/L,(75.65±0.33)U/L],第7天接近正常[(39.08±0.77)U/L,(62.23±0.29)U/L],但AST仍高于对照组。(2)HE染色:对照组结构正常,实验组肝小叶结构排列紊乱,气球样变明显,其中在12 h最为明显。(3)该方法操作简单,术中无明显副损伤,出血量极少,术后状态良好,二次术后存活率高(100%),造模成功率100%(30/30)。(4)本模型符合动物伦理要求。结论该肝损伤再生模型操作简单,易掌握,实用性强,成功率高,为研究肝再生、肝损伤、肝移植等提供了一种可靠且简便的动物模型。
Objective This study objective was to establish a model for repair and regeneration of acute liver injury in rats, featuring simple surgery and high survival rates. Methods A total of 30 Sprague-Dawley（SD） rats were randomly divided into a control group（n=6） and an experimental group（4 subgroups, 6 rats in each group, n=24）, and underwent surgery excising 70% of the left lateral lobe of the liver to simulate acute liver injury; in the control group, the venous blood of the trailing edge was examined to define the levels of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in normal liver tissues, and the surgery was immediately conducted to excise residual left hepatic lobe stained by hematoxylin-eosin staining（HE）, so as to clarify the normal liver tissue structure profile; in the experimental subgroups, their ALT and AST levels in venous blood of the trailing edge at the intervals of 6 hours, 12 hours, 3 days and 7 days were analyzed, and the surgery was immediately conducted to excise the residual left hepatic lobe stained by HE. The survival rate and the status of the experimental group were observed after two surgeries, with the successful modeling criteria being pathology standardization and survival in the long run. Results（1） Blood biochemical detection： compared with the control group, the levels of serum ALT and AST in the experimental group at the time points of 6 hours, 12 hours were significantly increased, and the difference was statistically significant（P0.05）, and at the time point of 6 hours, the levels were significantly increased [（200.150±1.2357） U/L,（257.483±1.1940） U/L], peaked at the time point of 12 hours [（449.933. 1.4949） U/L,（755.700±1.0040） U/L], began to decrease at the time point of 3 days [（55.87±0.88） U/L,（75.65±0.33） U/L], and closed to be normal at the time point of 7 days [（39.08±0.77） U/L,（62.23±0.29） U/L], but AST was still significantly higher than that of the control group.（2） HE staining： the liver structure of the control group was normal, while the hepatic lobules in the experimental group were arranged irregularly, and the balloon shape became more evident, peaking at the time point of 12 hours.（3） The surgery was performed with ease with no obvious side injuries, minimal bleeding, good postoperative conditions and high survival rates after two operations（100%）, and high success rate of molding（30/30）.（4） The model was consistent with animal ethical requirements. Conclusions The liver regeneration model has the advantages of being a simple surgical procedure, easy to master, very practical, and high success rates, it provides a reliable and simple animal model for the study of liver injury, regeneration and transplantation.
作者
王朝阳
朱美意
张宏伟
欧阳军
WANG Chaoyang;ZHU Meiyi;ZHANG Hongwei;OUYANG Jun(Department of general surgery;Department of Emergency Surgery;Department of Hepatobiliary Surgery,The First Affiliated Hospital of the Medical College,Shihezi University,Xinjiang Uygur Autonomous Region,Urumqi 832000,China)
出处
《中华灾害救援医学》
2017年第12期671-676,共6页
Chinese Journal of Disaster Medicine
基金
国家自然科学基金(81560312)
兵团应用基础研究等计划(2016AG019)
关键词
急性肝损伤
肝切除
肝再生
动物模型
大鼠
acute liver injury
hepatectomy
liver regeneration
animal model
rat
