D-氨基半乳糖诱导的猪急性肝功能衰竭模型的建立与改进

目的 建立和改进D-氨基半乳糖(D-gal)诱导的猪急性肝功能衰竭模型,探讨其应用于人工肝临床前评价的可行性.方法 19只杜洛克种猪分成4组,其中15只(每组5只)在无麻醉条件下通过颈静脉置管分别注射1.0、1.25及1.5 g/kg的D-gal,其余4只注射等量的5%葡萄糖液作为对照组.记录动物临床表现和存活时间,动态检测血氨、PT、肝肾功能、血糖、L-乳酸等指标和肝组织学变化.采用t检验及F检验比较上述指标的组间差异,采用Kaplan-Meier生存分析及LogRank检验比较动物存活时间.结果 注射D-gal 12 h后,所有动物出现明显的肝损伤表现,表现为血氨、AST、TBil及L-乳酸水平进行性升高,血糖水平显著下降,PT逐渐延长,与对照组比较,差异有统计学意义(F=32.33,F=27.817,F=50.097,F=88.382,F=8.211,F=21.227;均P＜0.01),尤以注射1.5 g/kg D-gal组的变化为著.接受1.0 g/kg D-gal的动物除2只存活超过168 h外,其余3只于68～84 h内死亡,而接受1.25 g/kg和1.5 g/kg D-gal的动物分别于33～89 h和23～47 h内死亡,死前均出现昏迷,病理学检查提示肝组织大片坏死伴出血.结论 在无麻醉条件下D-gal诱导的杜洛克种猪急性肝功能衰竭模型均具有潜在的可逆性和良好的重复性,且具有合适的治疗时间窗口,适宜于人工肝疗效和安全性评价研究.

Establishment and further improvement of D-galactosamine induced acute hepatic failure in Duroc breeding pigs

Objective To establish and improve the acute hepatic failure model in pigs induced with D-galactosamine (D-gal),and explore the feasibility of evaluating preclinical artificial liver devices.Methods Nineteen Duroc breeding pigs were divided into 4 groups.Fifteen unanesthetic Duroc breeding pigs out of 19 (5 of each group) received intravenously administration of D-gal at a dose of 1.0,1.25 and 1.5 g/kg body weight,respectively.The remaining 4 pigs which received the same volume of 5% dextrose in water served as controls. Clinical data and survival time of pigs were recorded.Blood samples were collected for dynamic testing of plasma ammonia,prothrombin time,liver and renal functions,blood glucose and L-lactate;liver tissues were sampled for pathological examination.The differences between groups were compared using t test and F test.The survival time of pigs was compared by Kaplan-Meier survival analysis and Log Rank test.Results Twelve hours after administration of D-gal,all pigs presented as acute hepatic failure characterized by progressive increases of levels of plasma ammonia,aspartate aminotransferase (AST),total bilirubin (TBil) and L-lactate,the level of blood glucose marked decreased and prothrombin time prolonged (F= 32.33,F=27.817,F=50.097,F=88.382,F=8.211,F=21.227;all P＜0.01);especially in the pigs which received D-gal at a dose of 1.5 g/kg.Except 2 pigs survived for 168 h,the other 3 pigs which received D-gal at a dose of 1.0 g/kg died within 68-84 h,while all pigs which received D-gal at a dose of 1.25 and 1.5 g/kg died within 33-89 h and 23-47 h,respectively.All pigs presented coma before death and liver histopathological examination indicated massive hepatic necrosis with severe hemorrhage.Conclusions D-gal induced acute hepatic failure model in unanesthetic Duroc breeding pig appears potential reversibility and high reproducibility,which has proper therapeutic window.Thus,this model could be applied to evaluate the therapeutic effects and safety of artificial liver devices.