采用免疫型肝硬化大鼠建立慢加急性肝衰竭模型方法的研究

目的对人血清白蛋白免疫诱导型肝硬化大鼠给予不同急性打击，探索建立与人慢加急性肝衰竭组织病理表现相似、实用性、重复性好的实验模型、方法用人血清向蛋白免疫诱导建立肝硬化大鼠模型，至肝纤维化达4级时随机分组，组一（n=6）给予1．2g／kg的D．氨基半乳糖腹腔注射；组二（n=6）给予30mg／kg脂多糖尾静脉注射；组i（n=6）给予D-氨基半乳糖400mg／kg＋脂多糖100trg／kg同时腹腔注射；组四（n：4）继续给人血清白蛋白静脉注射，计算每组动物自然生存时间及死亡率、观察各脏器组织病理变化。结果组一5只大鼠给药后39～52h之内死亡，肝组织病理显示肝硬化基础上发生弥漫大小泡脂肪变性，仅见小片坏死灶；组二大鼠均存活良好，给药后3天后处死肝组织未见坏死性病变。组三5只大鼠在13～19h内死亡，肝脏病理表现为再生结节大块或亚大块坏死，肝细胞凋广明硅，增生的纤维间隔完整保留。结论对人血自蛋白免疫诱导型肝硬化大鼠给予D-氨基半乳糖／脂多糖联合急性攻击可建立慢加急性肝衰竭模型，而单独给大剂量D-氨基半乳糖或脂多糖不能使肝硬化大鼠发生肝脏大块或亚大块坏死。

Exploration of establishing rat model of ACLF in human serum albumin-induced cirrhosis

Objective To establish a practical and reproducible animal model of acute-on-chronic liver failure, which is similar to the histopathology in human beings, we attacked the immunological cirrhosis rats with three means. Methods Immunological hepatic cirrhosis was induced by human serum albumin in Wistar rats. Rats with early-stage cirrhosis （fibrosis stage IV） were randomly divided into 4 groups. D-galactosamine （ 1.2 g/kg） was intraperitoneally administered in group 1 （n =6）. Rats in group 2 （n =6） were treated with lipopolysaccharide （30 mg/kg）. D-galactosamine （400 mg/kg） and lipopolysaccharide （100 ug/kg） were simultaneously injected in group 3 （n =10）, while group 4 （n = 4） were intravenous injected with human albumin as before. Survival time was observed, and liver injury was assessed by histopathology, as well kidney and lung. Results Hepatic fibrosis was detected after albumin intravenous injection for 6 weeks. Five rats died in group 1 after treatment with D-galactosamine, with survival time of 39 to 52 hours. Histopathology revealed severely adipose degeneration in regenerative nodules, only pelleta necrosis were seen. In group 2, rats experienced lethargy, febrile shaking and diarrhea, with fully recovery after 24 hours, three rats were sacrificed on 3 days, with no signs of necrosis in liver section. In group 3, 5 rats died from severe liver failure, with survival time of 13 - 19 hours. Histology of liver section revealed massive necrosis in nodules, with significant apoptosis of hepatocytes. Conclusion The experimental model of acute-on-chronic liver failure, induced by D-galactosamine/lipopolysaccharide in rats with immunological liver fibrosis, showed similar morphological changes as human beings, while D-galactosamine or lipopolysaccharide alone failed to sacrifice cirrhosis rats from liver massive necrosis.