大鼠腹腔注射人血清白蛋白免疫性肝纤维化模型的实验研究

Immune complex induced rat liver fibrosis model by intraperitoneal injection of human serum albumin

目的 通过腹腔注射人血清白蛋白建立免疫损伤性大鼠肝纤维化模型。方法 雄性Wistar大鼠，体重110—120g，皮下多点注射人血清白蛋白（共4次。分别间隔14、10、10d）。10d后，腹腔注射人血清白蛋白（每周2次，共8周，剂量从5mg逐渐增至20mg）。在攻击前、攻击后15、30、60d和停止攻击后30、60、90、120d，分别留取肝组织和血清标本，进行肝病理检查和肝羟脯氨酸生化测定，采用放射免疫法测定肝纤维化血清指标透明质酸（HA）和层粘连蛋白（LN）含量。结果 攻击后。肝纤维化分级、肝羟脯氨酸含量、血清HA、LN均升高（P〈0．05）。随造模时间延长，肝纤维化分级逐渐加重（P〈0．05）、肝羟脯氨酸含量、血清HA逐渐升高（P〈0．01）。停止造模后，肝羟脯氨酸含量、血清HA明显降低（P〈0．01），但仍显著高于正常对照组（P〈0．01）。纤维化形成率为100％，纤维化持续时间120d以上。结论 此造模方法简单，肝纤维化成功率高，维持时间长，可用于抗纤维化药物的研究。

Objective To establish an immune complex induced rat liver fibrosis model by intraperitoneal injection of human serum albumin （HSA）. Methods Male Wistar rats,weighting 110-120 g,were sensitized with HSA by subcutaneous injections at different sites for 4 shots at intervals of 14,10 and 10 days. Ten days after the fourth injection, the peritoneal booster dose of HSA was administrated to rats twice weekly for 8 weeks with an initial dose of 5 mg, and progressive increase to 20 mg. Liver biopsy was performed at the beginning of HSA booster, 15, 30,60 days after HSA booster, and 30,60,90,120 days after discontinuation of HSA booster, respectively. Liver samples were examined for histological changes and liver hydroxyproline （HyP） was measured by biochemical method. Fibrosis serum markers hyaluronate acid （HA） and laminin （LN） were determined by RIA method. Results After intraperitoneal administration of HSA, the degree of liver pathological changes, the liver Hyp content and serum HA and LN increased （ P 〈 0.05）. The longer the HSA administrated,the higher the liver pathological change degree （ P 〈 0.05）and the levels of liver Hyp and serum HA （ P 〈 0. 01 ）. After discontinuation of HSA, the levels of serum HA and liver Hyp decreased significantly （ P 〈 0.01 ） but were still significantly higher than those in the controls （P 〈 0. 01 ）. The liver fibrosis formation rate was 100% and the fibrosis lasted more than 120 days. Conclusions Intraperitoneal administration of HSA to make rat immune complex induced liver fibrosis model is convenient with high liver fibrosis formation rate and long fibrosis lasting time. The model may be used to evaluate the therapeutic effect of antifibrotic drugs.