Abstract: Objective To compare the characteristics of several modeling method and to find an optimal model for non-alcoholic fatty liver disease on the basis of ob / ob mice. Method Firstly, the second and third exons of leptin gene were knocked out in the background of C57 mice to obtain leptin knockout mice (B-ob / ob mice) , and then HFMCD induction, WD induction, WD combined with carbon tetrachloride induction were performed on the basis of B-ob / ob mice. The levels of ALT, AST, triglyceride and cholesterol in the serum of the mice were analyzed under these induction method; the NAS score of NAFLD was assessed by pathological HE staining; the degree of liver fibrosis was assessed by sirius red staining, and the degree of inflammation and fibrosis in the liver was also quantified by immunohistochemical staining. Result B-ob / ob mice had some fat accumulation and ballooning in the liver under normal dietary conditions without any induction. After 6 weeks of HFMCD induction, liver ALT was significantly elevated and liver fibrosis was increased. 12 weeks after WD induction, HE staining analysis showed some degree of intralobular inflammation in the liver in addition to steatosis and ballooning. After WD combined with carbon tetrachloride induction, HE staining showed that liver steatosis, ballooning and intralobular inflammatory features were more obvious, and liver fibrosis was also significantly increased, immunohistochemical result showed a significant increase in F4 / 80, a marker of hepatic macrophages, and α-SMA, a marker of hepatic stellate cell activation. Conclusion B-ob / ob induced by the combination of WD and CCL4 can mimic the pathological features of NAFLD more comprehensively in a short period of time, and is a better animal model of NASH.

Key words: ob / ob mice, non-alcoholic fatty liver disease, animal model

摘要： 目的 比较几种非酒精脂肪肝造模方法在 ob / ob 小鼠上的特征,建立基于 ob / ob 小鼠的非酒精性脂肪肝模型。 方法 首先在 C57 小鼠背景下,敲除瘦素基因的第二和第三个外显子,得到瘦素基因敲除的 ob 小鼠,即 B-ob /ob 小鼠,然后在 B-ob / ob 小鼠的基础上,分别进行HFMCD、WD 诱导、WD 联合四氯化碳诱导,并分析几种诱导方式下小鼠血清中 ALT、AST、三酰甘油和胆固醇的含量;病理 HE 染色进行非酒精性脂肪肝 NAS 积分评定;天狼星红染色进行肝纤维化程度评定,免疫组化染色对肝内炎性反应和纤维化程度进行定量分析。 结果 B-ob / ob 小鼠,在正常饮食条件下,不经任何诱导,肝中有一定的脂肪累积和气球样变。 在 HFMCD 诱导 6 周以后,肝 ALT 显著升高,肝纤维化增加。 WD 诱导 12 周以后,HE 染色分析除了脂肪样变和气球样变以外,肝出现一定程度的小叶内炎性反应。 WD 联合四氯化碳诱导后,HE 染色肝脂肪变,气球样变,小叶内炎性特征均比较明显,同时肝纤维化程度也明显增加,免疫组化结果显示肝巨噬细胞的标志物 F4 / 80 和肝星状细胞活化的标志物 α-SMA 都有显著的增加。 结论B-ob / ob 经过西方饮食和四氯化碳联合诱导,可以短时间内较为全面模拟非酒精性脂肪肝的病理特征,是一种较好的非酒精性脂肪肝病动物模型。

关键词: ob / ob 小鼠, 非酒精性脂肪肝, 动物模型