不同浓度四氯化碳诱导小鼠肝纤维化模型的比较

Comparison of Hepatic Fibrosis Model Induced by Different Concentrations of CCl_4 in Mice

目的对比观察不同剂量四氯化碳(CCl_4)诱导小鼠肝纤维化模型的效果,探索一种稳定可行且较为高效的肝纤维化小鼠造模方法。方法将60只C57BL/6J小鼠随机分为正常组和模型组,模型组小鼠分别腹腔注射体积分数10%的CCl4(5 mL/kg)以及体积分数20%的CCl_4(1 mL/kg),每周2次,连续8周。分别在造模2周、4周、6周、8周注射后处死小鼠,进行HE染色和Masson染色,检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,以及肝匀浆氧化/抗氧化指标,丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH)和超氧化物岐化酶(SOD)变化,Western blotting法检测肝组织中胶原表达情况。结果随着造模时间的延长,病理染色显示肝纤维化程度不断加重,胶原表达逐步增加,血清ALT、AST水平逐渐升高,肝匀浆SOD、GSH活性降低,氧化产物MDA含量升高。在造模的相同时间点,体积分数10%CCl_4、5 m L/kg剂量组相比于体积分数20%CCl4、1 m L/kg剂量组,病理染色显示肝纤维化程度更加严重,血清ALT、AST水平明显升高,肝脏中SOD、GSH活性明显降低,氧化产物MDA含量明显升高,肝纤维化形成的时间更短,效果更好。结论两种造模方法都能形成肝纤维化小鼠模型,但体积分数10%CCl_4(5 m L/kg)剂量组的纤维化形成时间更短且效果更好。因此,每周2次,连续8周腹腔注射10%CCl_4(5 m L/kg)可作为一种较为理想的纤维化小鼠造模方法。

Objective To explore a reliable and stable method for establishing hepatic fibrosis model in mice through different dosing level of carbon tetrachloride（CCl4）. Methods Sixty male C57 BL/6 J mice were randomly divided into control group and model group. The mice in model groups were intraperitoneally injected with 10% CCl4 at the dosage of 5 m L/kg and 20% CCl4 at the dosage of 1 m L/kg respectively for 8 weeks, twice per week. Mice were sacrificed at the 2 nd, 4 th, 6 th, 8 th week after injection. The degree of hepatic fibrosis was evaluated by HE and Masson staining. The changes of alanine aminotransferase（ALT）, aspartate aminotransferase（AST） were detected in serum. The oxidative index malondialdehyde（MDA） and antioxidant indices, glutathione（GSH）, superoxide dismutase（SOD） were detected in liver homogenate. The protein levels of collagen I and collagen III in liver tissues were determined by Western blotting. Results HE and Masson staining showed that the degree of hepatic fibrosis was aggravated as the development of the model. The expression of collagen was gradually increased, the serum levels of ALT, AST and MDA in liver tissues were significantly increased, while the activities of SOD and GSH were significantly decreased. At the same point of model progress, the degree of hepatic fibrosis of 10% CCl4（5 m L/kg） group was more severe than that of 20%（1 m L/kg）group. The levels of ALT, AST and MDA in 10% CCl4（5 m L/kg） were significantly higher than 20%CCl4（1 m L/kg） group. In addition, the SOD activity and GSH were lower than those of 20%（1 m L/kg）group. Conclusion Both of two methods can induce hepatic fibrosis model, and 10% CCl4（5 m L/kg）has advantages such as better effect and shorter modeling time compared with 20% CCl4（1 m L/kg）,therefor, it is an ideal method for establishment of liver fibrosis model in mice.