四氯化碳诱导大鼠肝硬化腹水模型的改进与优化

Improvement and optimization of CCl_4 induced liver cirrhosis rats model accompanied with ascites

目的应用四氯化碳(CCl4)联合苯巴比妥及乙醇的方法,以期建立稳定性、均一性、重复性良好且成模率较高的大鼠肝硬化腹水模型,为相关血浆蛋白制品的药效学评价等研究奠定基础。方法将90只雄性Wistar大鼠随机分为5组:正常对照组10只,腹腔注射橄榄油,正常饮水;模型组分为CCl4组、CCl4+苯巴比妥组、CCl4+乙醇组、CCl4+苯巴比妥+乙醇组,每组20只,给予不同饮用水处理,腹腔注射40%CCl4橄榄油混合液。各实验组腹腔注射剂量均为2 ml/kg,每周注射2次,共12周。建模过程中,定期测量体质量和腹围,采集静脉血监测肝功能指标和血浆胶体渗透压(COP)。建模结束后,随机处死各组大鼠,取肝右叶组织进行病理学鉴定。应用统计学方法对数据进行分析。结果各模型组在建模的第8～12周先后出现符合筛选标准的模型。其中,第8周时,CCl4+苯巴比妥+乙醇组大鼠开始出现腹水阳性体征,个别达到建模标准;第9周时,CCl4+苯巴比妥组和CCl4+乙醇组均有符合要求的模型产生;CCl4组至第10周时开始出现建模成功的大鼠。至12周建模结束时,与正常对照组相比,各模型组体质量、腹围、肝功能指标、COP等差异均具有统计学意义,以CCl4+苯巴比妥+乙醇组的差异最为明显。各模型组的成模率分别是65%、75%、75%和80%。镜下观察模型组大鼠的肝组织病理学切片,可见假小叶形成等典型的肝硬化形成特征。结论四氯化碳联合苯巴比妥与乙醇诱导大鼠肝硬化腹水动物模型较传统的方法可以有效提高实验动物的成模率,缩短建模周期。

Objective To establish a stable, homogeneous, well-tolerated and efficient protocol to induce rat cirrhosis models accompanied with ascites using an improved carbon tetrachloride（ CC14 ） combined with phenobarbital sodium and alcohol method, thus establishing a basis for serum albumin effectiveness evaluation test. Methods Ninety male Wistar rats were randomly divided into five groups ： blank control group containing 10 rats, CC14 group, CC14 ＋ phenobarbital sodium group, CC14 ＋ alcohol group and CC14 ＋ phe- nobarbital sodium ＋ alcohol group containing 20 rats respectively as model groups. The control group was given normal drinking water and injected with olive oil intraperitoneally. Rats in model groups were given different drinking water and an intraperitoneal injection of CC14 in a 2：3 mixture with olive oil. The drugs given intraperitoneally were all at a dose of 2 ml/kg body mass, twice weekly for 12 weeks. We further measured the body mass and abdominal circumference, and tested the biochemical liver function indices as well as the colloid osmotic pressure （COP） to evaluate the different processes in these groups. At the end of the induced operation the random- ly selected rats in each group were sacrificed, and a section of right liver lobe was obtained and examined by histopathological method. The statistical analysis data were performed. Results Cirrhosis accompanied with ascites developed in all model groups between 8 and 12 weeks. However, at the end of the eighth week, rats in CC14 ＋ phenobarbital sodium ＋ alcohol group were firstly detected to corre- spond with the model standards, CC14 ＋ phenobarbital sodium group and CC14 ＋ alcohol group were detected to have positive model signs at the ninth week. Only at the tenth week a few rats appeared to have aseites in CC14 group. All the model groups showed signifi- cant differences in the biochemical liver function indices, COP and body mass than the rats in control group, and the most significant differences were observed in CC14 ＋ phenobarbital sodium ＋ alcohol group. The model rate in model groups was 65%, 75%, 75% and 80% . resnectivelv. Histological examination showed that all the model groups developed liver cirrhosis with typical pseudolobule forma-tion. Conclusion The use of carbon tetrachloride associated with phenobarbital sodium and alcohol presents an effective method to in- duce cirrhosis rat models with ascites, the model rate has a good advantage than other transitional methods.