摘要： 目的 建立幼鼠的非酒精性脂肪性肝炎（NASH）模型，并探讨幼鼠建模中肝纤维化相关的胶原参数动态变化特点，为研究儿童NASH肝纤维化的进展提供新思路。方法 3周龄、4周龄、5周龄和6周龄小鼠喂养蛋氨酸胆碱缺乏饲料（MCD），并设立相应的对照组喂养普通饲料，共喂养8周。在喂养的0、4、6和8周时采集小鼠肝组织，行HE和天狼猩红染色，观察肝组织病理特征并评分。同时对肝组织进行二次谐波（SHG）/双光子激发荧光（TPEF）显微成像，定量分析100个胶原参数，以造模后不同时间点和不同纤维化分期为标准，分析不同周龄小鼠的胶原特征。结果 与对照组相比，随着MCD造模时间的延长，不同周龄的小鼠体质量逐渐下降，幼鼠死亡数量增加，丙氨酸氨基转氨酶（ALT）和天冬氨酸氨基转氨酶（AST）升高。MCD诱导4周后，各组小鼠的肝脏出现不同程度的脂肪变和炎症，诱导8周后，胶原含量明显增加。3、4、5周龄小鼠的汇管区胶原含量高于6周龄小鼠，中央静脉区和窦周区胶原含量均低于6周龄小鼠。各组16个胶原总特征的指标无显著差异。结论 3、4、5周龄幼鼠的NASH模型以汇管区胶原变化为主，病理改变更符合儿童NASH病理特征，适合建立儿童脂肪肝模型。

幼鼠非酒精性脂肪性肝炎模型的建立及其肝纤维化特征的研究

武楠¹, 王晓晓¹, 费然¹, 饶慧瑛¹, 魏来², 刘峰¹

1. 北京大学人民医院, 北京大学肝病研究所, 丙型肝炎和肝病免疫治疗北京市重点实验室, 北京 100044;
2. 清华大学附属北京长庚医院肝胆胰中心, 北京 102218

收稿日期:2019-12-03出版日期:2020-03-30发布日期:2020-03-19
通讯作者:刘峰E-mail:liu1116m@sina.com
作者简介:武楠(1980-),女,主治医师,博士.
基金资助:政府间国际科技创新合作重点专项（中国与新加坡双边合作项目）（2016YFE0116800）；"重大新药创制"科技重大专项（2018ZX09201002）；北京大学人民医院研究与发展基金（RDX2018-06）


关键词: 非酒精性脂肪性肝炎, 幼龄, 胶原定量, 肝纤维化
Abstract: Objective To establish a juvenile nonalcoholic steatohepatitis (NASH) model and observe the progression of liver fibrosis. Methods Mice of different ages (3-,4-,5-,and 6-week-old) were induced with methionine-and choline-deficient diet (MCD). Serum and liver tissue specimens were collected at 0,4,6,and 8 weeks after induction. HE and Sirius red staining were performed for histological assessment. SHG/TPEF imaging was performed and 100 collagen parameters,including those for the overall (16 parameters),portal (28 parameters),central vein (28 parameters),and perisinusoidal (28 parameters) collagen in liver tissue,were quantified. The time-based progression of fibrosis was analyzed. Results The MCD diet gradually decreased the body weight of mice at different weeks,compared to that in the control group. It increased the mortality and alanine aminotransferase (ALT) and aspartate transaminase (AST) levels in mice at 4 weeks. Mice of different ages showed varying degrees of hepatic steatosis and inflammation. After 8 weeks of induction with MCD,the collagen content was observed to increase significantly. Compared to 6-week-old mice,3-,4-,and 5-week-old mice had higher content of periportal collagen,and lower content of central vein and perisinusoidal collagen. There were no differences in the 16 parameters determined for collagen over all the regions among mice of all the ages. Conclusion Changes in the collagen present in the portal area were observed in 3-,4-,and 5-week-old mice with NASH. These mice would be suitable as a model of pediatric NASH.
Key words: nonalcoholic steatohepatitis, juvenile, collagen quantification, liver fibrosis
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