大鼠门静脉分支残端置管模型构建及细胞移植途径的评价

Establishing a Model of Acute Hepatic Failure with Catheterization of Portal venous Branch and a Study of the Way of Cell Transplantation

目的:大鼠肝部分切除模型被广泛的应用于肝脏疾病的研究,随着干细胞治疗肝损伤及护肝药物研究的发展,对大鼠肝损伤模型也提出了很多新的要求。本实验拟在大鼠肝部分切除术的基础上改进以建立大鼠肝断面门静脉分支残端的静脉置管模型,并进行细胞移植实验,对比分析新模型的优劣。方法:60只F344大鼠分为三组。A、B组行行85%肝切除术;C组行85%肝切除术+肝断面门静脉分支残端置管术。术中B组经门静脉注入4×105个表达GFP(green fluorescence protein,GFP)的胎肝干细胞(fetal liver stem/progenitor cells,FLSPCs)。C组经留置导管注射入同等量的FLSPCs,A组注射同等剂量的培养液。72小时取血清,测定肝功能ALT、AST,统计死亡率;取肝脏组织切片观察其修复情况。统计学采用方差分析和LSD-t检验。结果:B、C组F344大鼠72小时肝功指标(ALT、AST)均明显优于A组;B组、C组肝脏组织学的病理损伤的恢复分别较A组快。B、C组间肝功指标无统计学意义。结论:经门静脉分支残端置管途径移植FLSPCs效果等同于经门静脉穿刺途径,且该模型具有可反复、可选时、减少创伤等优点。

Objective： Rat partial hepatectomy model is widely used in the study of hepatic disease. As the rapid development of research of liver protection drugs and stem cells therapy for liver injury, the model need to be improved. A rat model of acute hepatic failure with catheterization of portal venous branch was established. After a study of the way of cell transplantation was made in our trail, the new model was evaluated. Methods： Sixty F344 rats were randomly divided into 3 groups （n=20 in each group）. The partial hepatectomy with 85% sizes were performed on the rats of the A and B groups. The same operations and catheterizations of portal venous branch were carried out on the rats of the C group. After the green fluorescent protein （GFP） gene was transferred into the FLSPCs, Every rats of the B group were transplanted into 4×105 cells through the portal veins during the operations. Every rats of the C group were transplanted into the same cells through the ducts. Control rats in the A group received medium of equal volumes. Mortality rates and hepatic functions such as ALT AST were measured after 72 hours. Meanwhile, hepatic pathology was studied. Data were analyzed using variance analysis and LSD-t. Results： Compared to control group the hepatic and coagulative functions gradually recovered in stem cells transplant groups（B and C）. The hepatic pathology significantly improved in stem cells transplantation groups（B and C）. The effects of treatments on the rats of B group are equal with those of C group. Conclusions： Stem cells transplantations through ducts of portal venous branch improved the hepatic ftmctions and pathology in acute hepatic injury model of F344 rats as much as those through portal vein directly. The model has advantages in repeated operation, fewer traumas and less complication and can be treated whenever necessary.