门静脉注射D-氨基半乳糖和脂多糖建立猪急性肝功能衰竭模型

Establishment of a porcine model of acute hepatic failure by intraportal injection of D-galactosamine and lipopolysaccharide

目的通过门静脉注射D-氨基半乳糖(DGal)和脂多糖的方法建立一种接近临床条件的猪急性肝功能衰竭(AHF)模型。方法22头健康家猪随机分为5组。组Ⅰ:门静脉注射生理盐水(3头);组Ⅱ:门静脉注射1μg/kg脂多糖(5例);组Ⅲ:门静脉注射0.5g/kgD-Gal(5例);组Ⅳ:门静脉注射0.5g/kgD-Gal+1μg/kg脂多糖(6例);组Ⅴ:门静脉注射0.5g/kgDGal+1μg/kg脂多糖,接受辅助性部分原位肝移植术(APOLT)(3例)。观察各组动物的临床表现、生化指标和病理改变。结果组Ⅰ和组Ⅱ动物全部存活,生化指标轻度改变。组Ⅲ动物1周内死亡率为40%(2/5)。组Ⅳ中5例(5/6,83%)在120h内死亡,注药后48h血清天冬氨酸氨基转移酶达最高峰(4912U/L±759U/L),同时伴有总胆红素、乳酸、血氨明显升高和凝血酶原时间延长。组Ⅴ动物行APOLT治疗后AHF逆转,第3天起各生化指标开始恢复,至第7天基本恢复正常,病理结果显示原肝得到部分再生。结论通过门静脉注射0.5g/kgDGal和1μg/kg脂多糖的方法可建立一种理想的猪急性肝功能衰竭模型。该模型可用于辅助性部分原位肝移植治疗急性肝功能衰竭的研究。

ObjectiveTo develop a clinically relevant porcine model of acute hepatic failure (AHF). MethodsTwenty-two healthy pigs were randomly divided into 5 groups: group Ⅰ(n=3, intraportally administered with normal saline), group Ⅱ [n=5, intraportally administered with 1 μg/kg of lipopolysaccharide (LPS)], group Ⅲ [n=5, intraportally administered with 0.5 g/kg of D-galactosamine (D-Gal)], group Ⅳ (n=6, intraportally administered with 0.5 g/kg of D-Gal plus 1 μg/kg LPS), and group Ⅴ [n=3, intraportally administered with 0.5 g/kg of D-Gal plus 1 μg/kg LPS and then receiving auxiliary partial orthotopic liver transplantation (APOLT)], Blood samples were collected to examine the arpartate transaminase (AST), total bilirubin, lactic acid, blood ammonia, prothrombin time (PT), blood sugar, and creatine at different time points. Autopsy was performed on the dead animals. Eight days after the APOLT laparotomy was performed again on the surviving pigs to take specimens of the original and transplanted liver to undergo pathological examination. ResultsAll the pigs in the groupsⅠand Ⅱ survived with minimal changes in liver function tests. Two of the 5 pigs in the group Ⅲ died (40%), 5 pigs in the group Ⅳ (5/6,83%) died within 120 h, with a significant increase in aspartate transaminase 48 h after (4912 U/L±759 U/L). In comparison with those of the group 1 and 2, the TBIL, blood ammonia, lactic acid, and PT 48 h after of the group 4 were all significantly higher and the blood sugar was significantly lower (all P<0.05). Reversal of AHF in the pigs in the group Ⅴ following APOLT was observed and the liver function returned near to normal level on the 7 th postoperative day and regeneration of the native liver was confirmed histologically. ConclusionThe porcine model of AHF induced by a combination of D-gal ( 0.5 g/kg) and LPS (1 μg/kg) will be of much use in the development of APOLT for AHF.