冷保存、慢性排斥反应构建移植肝胆管病大鼠模型  

The rat model of graft cholangiopathies established by cold preservation or chronic rejection

作　　者：陆宏伟[1,2] 张玉君[3] 余强[2] 李崇辉[2] 

机构地区：[1]西安交通大学第二附属医院普通外科,710004 [2]解放军总医院肝胆外科,北京 [3]第三军医大学西南医院肝胆外科,重庆

出　　处：《中华肝胆外科杂志》2013年第8期601-605,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：基金项目：国家自然科学基金（81170454）

摘　　要：目的建立大鼠移植肝胆管病模型并评估其意义。方法将大鼠按实验要求分为4组：（1）长时间（12h）冷保存组（n=24）：同基因系近交系大鼠♂Wistar→♂Wistar，所获供肝于4℃UW液保存12h后行两袖套法大鼠原位肝移植（rat liverorthotopictransplantation，ROLT）。术中利用内支架管直接对合供、受体肝总动脉及肝外胆管；（2）慢性排斥反应组（低剂量环孢素每天1mg／kg，冷缺血时间1h）（n=24）：异基因近交系大鼠♂DA→♂Lewis。供肝于4℃UE液保存1h后同法行ROLT；（3）对照组（冷保存时间1h）（n=24）：同基因系近交系大鼠♂Wistar→♂Wistar，供肝于4℃UW液保存1h后同法行ROLT；（4）假手术组（n=24）：♂Wistar大鼠，只进行开关腹手术。观察术后16周内各组大鼠并发症的发生情况和大鼠肝脏标本的病理学改变。结果在长时间冷保存组和慢性排斥反应组，术后大鼠意识恢复慢，胆道及血管并发症发生率高，肝内汇管区胆管增生及炎细胞浸润明显。16周时可见肝小叶被增生胆管分隔，正常肝小叶结构消失，增生胆管上皮细胞萎缩、坏死、胞浆消失，仅见固缩的细胞核。胆管周围有较多的炎细胞浸润，肝内小动脉闭塞或消失。结论通过供肝长时间冷保存或诱导慢性排斥反应可以建立稳定可靠的大鼠移植肝胆管病模型。该模型是研究肝移植术后移植肝胆管病的理想模型。Objective To develop rat models of graft cholangiopathies and evaluated their values. Methods Four groups were constructed. The long-term cold preservation group（n= 24） contained homogeneic inbred rat liver orthotopic transplantations （ROLT） performed in a rat combination of ♂ Wistar→♂ Wistar with the donor liver preserved in 4℃ UW for 12 h, the vessels reconstructed by the two-cuff method, and the hepatic artery and extrahepatic bile duct rebuilt by a stent. The chronic rejection group （n=24）（CsA 1 mg . kg^-1 . d^-1, cold preserved for 1 h） was allogeneic inbred DA→♂ Lewis rats induced for ROLT, and the revascularization methods were the same as the longterm cold preservation group. The control group （n=24） （cold preserved for 1 h） was homogeneic inbred ♂ Wistar→♂ Wistar rats with ROLT techniques the same as above. The sham group （n= 24） was ♂ Wistar rats that had an exploratory laparotomy. The animals were followed for 16 weeks, complications were compared, and liver tissues were harvested. Histopathological and morphometric techniques were used to construct a time course of histological changes after liver transplantation. Results In both the long-term cold preservation group and chronic rejection group, the rats recovered slowly, the incidence of complications and mortality were higher than those of other groups, and the intrahepatic bile duct proliferation and immune cell infiltration were noticeable after the operation. In 16 weeks, the hepatic lobules were separated by the proliferating bile ducts, the normal structure of hepatic lobules disappeared, many biliary epithelial cells necrosed with disappearing cytoplasms, and there was immune cell infiltration and obliterative arteriopathy. Conclusions The rat models of graft cholangiopathies induced by long-term cold preservation or chronic rejection donor livers are stable and easily standardized. This model is ideal for studying the pathogenesis and prevention of graft cholangiopathies.

关 键 词：大鼠模型 移植肝胆管病 冷保存 慢性排斥反应 

分 类 号：R-332[医药卫生]