摘要
目的建立一个模拟人类疾病进程的猪急性肝衰竭（ALF）模型，用于ALF治疗药物的临床前安全性评价及疗效评价；检测模型中猪纤维介素（pfgl2）的表达情况，为针对垃12的基因治疗提供基础和依据。方法造模组耳静脉陕速注射D-氨基半乳糖盐酸盐，剂量1．2g／kg；对照组耳静脉快速注射5％的葡萄糖，剂量12ml／kg。观察两组动物临床表现、肝功能指标和肝组织病理学改变；实时定量PCR检测肝组织中pfgl2 mRNA表达，免疫组织化学检测肝组织中pfgl2蛋白的表达。采用重复测量数据的方差分析和独立样本t检验进行统计学处理。结果成功建立了与人在临床表现、肝脏生物化学指标、组织病理学改变相似的猪ALF模型；实时定量PCR检测结果显示造模组猪肝组织中pfgl2的mRNA表达水平显著增加，与对照组比较，差异具有统计学意义（t=7．695，P〈0．05）。免疫组织化学显示造模组猪肝组织中有明显的pfgl2蛋白的表达，主要分布在肝细胞坏死区域的肝细胞、炎症浸润细胞、肝血窦内皮细胞及血管内皮细胞，对照组动物肝组织未见pfgl2阳性着色。结论以D-氨基半乳糖盐酸盐诱导的猪ALF模型可用于评价肝衰竭治疗药物的临床前疗效及安全性；pfgl2在猪ALF动物模型的肝组织中异常高表达，提示其参与了ALF时肝细胞坏死的发生和发展过程。
Objective To establish a pig model of fulminant hepatic failure for evaluating the preclinical efficacy of drug treatment on severe hepatitis, and to detect the expression of fibrinogen-like protein- 2 （fgl2） prothrombinase in the model, so as to provide basis for gene therapy targeting to fgl2 for fulminant hepatic failure. Method D-galactosamine hydrochloride was used to induce pig model of fulminant hepatic failure, and the experiment animals were divided into model group （rapid injection of D-galactosamine hydrochloride by ear vein, a dose of 1.2 g/kg） and negative control group （5% Glucose）. Clinical, biochemical and pathological changes of animals were observed. The expression of pigs fgl2 （pfgl2） mRNA in liver tissue was detected by real time RT-PCR, the expression of pfgl2 protein in liver tissue was detected by immunohistochemistry. Results A pig model of fulminant hepatic failure was successfully established using the D-galactose hydrochloride; Real time RT-PCR of liver fgl2 mRNA showed that fgl2 mRNA expression was increased significantly in liver tissue of fulminant hepatic failure pig model compared with the control group （P = 0.016）; Immunohistochemical staining showed that there were fgl2 protein expression in liver tissue of fulminant hepatic failure pig model, mainly in the membrane and cytoplasm of hepatocytes, inflammatory cells, liver sinusoidal endothelial cells and vascular endothelial cells of liver cell necrosis region.However, there are no fgl2 positive staining on negative control. Conclusions The pig model of fulminant hepatic failure induced by D-galactosamine hydrochloride is similar to human pathological process and can be used to evaluate the pre-clinical efficacy and safety of drug treatment on fulminant hepatic failure. Abnormal expression of pfgl2 at both mRNA level and protein level in the liver of fulminant hepatic failure pig model shows that pfgl2 induced coagulation pathway is also involved in the development of fulmiuant hepatic failure. Gene therapy targeting fgl2 genes for fulminant hepatic failure may provide a new means for the treatment of fulminant hepatic failure.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2009年第9期691-694,共4页
Chinese Journal of Hepatology
基金
国家重点基础研究发展计划（973计划）（2007CB512900）
“十一五”国家科技支撑计划（2006BA105A07）
关键词
肝功能衰竭
急性
动物模型
猪纤维介素
Liver failure, actue
Animal model
Pig fibrinogen-like protein 2
