钳夹法建立小鼠胆管缺血狭窄模型

目的 探讨建立小鼠胆管缺血狭窄模型的方法。方法 先探索昆明小白鼠胆管缺血最适时间点，尔后选取昆明小白鼠随机分为正常对照组和模型组，每组10只，模型组用2枚显微血管夹（宽0．1cm）夹闭一段长约0．4cm的胆总管90min，正常对照组仅暴露胆管未夹闭。术后21d两组小鼠均行胆道造影，下腔静脉取血，分离血清进行总胆红素及谷丙转氨酶含量测定，取胆总管和肝组织石蜡包埋，HE染色，观察病理改变。结果 模型组钳夹部位以上胆管明显扩张，胆管造影示胆管扩张，壁光滑，狭窄部位以下胆管未显影；正常对照组肝内胆管未显影，肝外胆管显影。血清总胆红素模型组高于正常对照组5倍以上，谷丙转氨酶模型组显著高于正常对照组6倍以上。病理示缺血胆管管腔变小或闭塞，胆管黏膜坏死脱落，管壁纤维化增厚；病变部位以上胆管管腔增大，管壁变薄。肝脏可见少量肝细胞坏死，炎性细胞浸润。结论 血管夹钳闭缺血法成功地建立了小鼠胆管缺血狭窄模型，为肝移植术后胆管缺血狭窄并发症发生机制的实验研究和临床研究提供可靠的实验方法。

Establishment of an animal model of biliary ischemic stenosis with clamping in mice

Objective To explore the method for establishing biliary ischemic stenosis model in mice.Methods After the optimal time point of biliary ischemia was determined,20 Kunming mice were equally divided into 2 groups.A 0.4 cm length of common bile duct was clamped for 90 min with 2 micro-vessel clamps(width=0.1 cm)in the experimental group.The common bile duct was not clamped in the control group.Twenty-one days later,biliary tract visualization was conducted in all the mice.Blood samples were collected form the inferior vena cava to determine the serum levels of TBIL and ALT.Meanwhile,the samples of the common bile duct and liver tissues were extracted for microscopic examination to observe the morphological changes.Results In the experimental group, obvious dilatation of the common bile duct appeared over the clamped site.There was no dilatation of the common bile duct in the control group.Twenty-one days inter,the serum levels of TBIL and ALT were significantly higher in the experimental group than in the control group.Microscopic examination showed that the part of common bile duct over the clamped site significantly expanded,the bile duct lumen became smaller or occluded,mucosa of the bile duct had necrosis and fell off and tubular wall had fibrosis and incrassation.A few dead liver cells and a lot of inflammatory cells were seen in the samples of liver tissues.Conclusions The biliary ischemic stenosis model has been successfully estab- lished with the clamping method in mice and it may provide a reliable experimental method for basic and clinical researches of mechanism of biliary ischemic stenosis after liver transplantation.