四氯化碳致肝纤维化动物模型实验条件的优化

Optimization of an experimental model of hepatic fibrosis induced by carbon tetrachloride

目的优化四氯化碳(carbon tetrachloride,CCl4)肝纤维化动物模型的造模方法。方法对肝纤维化模型从CCl4剂量、造模途径和动物种属等方面进行优化。造模剂量优化时采用昆明小鼠,腹腔注射CCl40.5、0.75、1.0ml/kg,2次/周,共8周,于第9周开始停止注射CCl4,观察至第12周;造模方式优化时采用ICR小鼠,CCl4腹腔注射或灌胃,2次/周,共12周;动物种属优化时,选择ICR小鼠和SD大鼠,CCl4灌胃,2次/周,共12周。行苏木精-伊红染色和马松三色染色观察动物的肝脏病理学改变。结果选用不同剂量的CCl4腹腔注射后,在第10周,0.5ml/kgCCl4造模仅在汇管区形成纤维化,0.75ml/kgCCl4则形成了纤维间隔,而1ml/kgCCl4造模可以形成早期肝硬化,提示适当的CCl4剂量为1ml/kg。灌胃和腹腔注射的方式均能导致明显的肝纤维化,其中灌胃方式死亡率在10%以下,而腹腔注射方式造模死亡率在80%以上,提示灌胃造模动物耐受性较好。应用CCl4后,大鼠和小鼠肝纤维化的各指标变化趋势一致,大鼠变化更为明显。结论 CCl4致肝纤维化模型的建立宜采用1ml/kgCCl4灌胃的方法,可根据实验目的选用不同种属的动物。

Objective To optimize the liver fibrotic animal model induced by carbon tetrachloride （CCl4）.Methods The optimization of the liver fibrotic animal model involved doses of CCl4,routes of CCl4 administration and animal species.To find the suitable dose of CCl4 for modeling,KM mice were injected intraperitoneally with CCl4 at the dose of 0.5,0.75,1 ml/kg body weight,twice a week,for a total of 8 weeks.From the beginning of the 9th week CCl4 injection was stopped,but the test lasted until the 12th week.To optimize the routes of CCl4 administration,ICR mice were treated with CCl4 intraperitoneally or intragastrically,twice a week,totally 12 weeks.To compare mice and rats for modeling,we administrated CCl4 to ICR mice or SD rats intragastrically,twice a week,totally 12 weeks.Results After 8 weeks of CCl4 injection,in the 10th week there were collagen fibers limited in portal areas of the 0.5 ml/kg CCl4 group,fibrous septum was formed in the 0.75 ml/kg CCl4 group and early cirrhosis in the 1 ml/kg CCl4 group,suggesting that the appropriate CCl4 dose was 1 ml/kg.Although intraperitoneal injection and intragastric administration both induced apparent fibrosis,the death rate after intraperitoneal injection was up to 80%,vs 10% or below after intragastric administration which proved to be a better administration.Trends of liver fibrotic indicators of rats and mice were comparable and changes of rats were more evident.Conclusion To establish liver fibrosis induced by CCl4,1 ml/kg CCl4 and intragastric administration should be adopted and animal species should be chosen according to the purposes.