三种非酒精性脂肪肝炎模型病证特点的比较研究  

Comparison research of disease characteristics in three non-alcohol steatohepatitis models

作　　者：薛静波 杨金凤 黄恺 彭渊 陶艳艳 刘成海 XUE Jingbo;YANG Jinfeng;HUANG Kai;PENG Yuan;TAO Yanyan;LIU Chenghai(Institute of Liver Diseases,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Key Laboratory of Traditional Chinese Clinical Medicine,Shanghai 201203,China;Key Laboratory of Liver and Kidney Diseases(Ministry of Education),Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肝病研究所,上海201203 [2]上海市中医临床重点实验室,上海201203 [3]上海中医药大学肝肾疾病病证教育部重点实验室,上海201203

出　　处：《中国实验动物学报》2025年第1期34-43,共10页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金面上项目(82274305);上海市临床重点专科建设项目(shslczdzk01201)。

摘　　要：目的比较四氯化碳(carbon tetrachloride,CCl_(4))注射复合高脂饲料饮食(high fat diet,HFD)、蛋氨酸胆碱缺乏饮食(methionine and choline deficient diet,MCD)以及高脂高糖高胆固醇饮食(aymlin liver NASH,AMLN)诱导的三种非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)模型血清学及病理学特征。方法分别采用CCl_(4)注射复合高脂饲料饮食(CCl_(4)+HFD)、MCD、AMLN喂养10、8和6周,制备小鼠NASH模型。比色法检测血清丙氨酸转移酶(alanine aminotransferase,ALT)、天门冬氨酸转移酶(aspartate aminotransferase,AST)和血糖(glucose,GLU)的水平及肝组织甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)和丙二醛(malondialdehyde,MDA)含量及超氧化物歧化酶(superoxide dismutase,SOD)活性;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清胰岛素(fasting insulin,FINS)含量并计算胰岛素抵抗指数(homeostasis model assessment of insulin resistant,HOMA-IR);苏木素-伊红(hematoxylin-eosin,HE)染色、天狼猩红染色及油红O染色观察肝组织炎症、脂滴生成及胶原沉积情况,并根据非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)活动度评分(NAFLD activity score,NAS)对肝组织脂肪性肝炎分级评分。结果与各正常组小鼠相比,3种模型组小鼠血清ALT、AST活性、肝TG、TC、MDA含量及SOD活性均有显著上升;其中AMLN模型组及CCl_(4)+HFD模型组小鼠的血清FINS、GLU的含量显著升高,HOMA-IR指数显著升高;MCD模型组小鼠血清FINS、GLU的含量及HOMA-IR指数显著下降。HE染色、油红O染色及NAS评分结果显示,3种模型组小鼠的肝组织均已进展到脂肪性肝炎阶段,且CCl_(4)+HFD模型组小鼠肝组织胶原沉积最明显,AMLN模型组小鼠肝脂滴最丰富。结论上述三种模型均可稳定模拟人类NASH疾病的血清学及病理学变化,其中AMLN模型可模拟人类的发病过程及机制,且胰岛素抵抗、氧化应激等全身代谢紊乱表现,Objective To compare the serological and pathological characteristics of 3 nonalcoholic steatohepatitis(NASH)models:high-fat diet(HFD)with carbon tetrachloride(CCl_(4))injection,methionine and choline deficient diet(MCD),and Aymlin liver NASH(AMLN)diet-induced NASH models.Methods 3 NASH models were established by feeding mice an HFD with CCl_(4)injection for 10 weeks,MCD for 8 weeks and NASH for 26 weeks.After feeding,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),glucose(GLU),liver triglyceride(TG),total cholesterol(TC),and malondialdehyde(MDA)levels and superoxide dismutase(SOD)activity were measured.Insulin levels were measured by enzyme-linked immunosorbent assay(ELISA)and the homeostasis model assessment of insulin resistant(HOMA-IR)index was calculated.Hematoxylin-eosin(HE),Sirius red,and oil red staining were used to indicate pathological changes to the liver.The NAS score was used to grade the pathology.Results Compared to each normal control(NC)group mice,all mice in the 3 model groups had an obvious increase in serum transaminase and liver TG,TC,MDA levels and SOD activity.The levels of serum FINS,GLU and the HOMA-IR index were significantly increased in the AMLN and CCl_(4)+HFD model groups but decreased in the MCD model group.According to the HE,oil red staining and NAS score,mice in all 3 groups had NASH phenotypic changes.Liver collagen deposition was most obvious in mice in theCCl_(4)+HFD model group.Liver lipid droplets were most abundant in the AMLN model group.Conclusions All the above 3 animal models can stably simulate the serological and pathological changes of NASH in human.The AMLN model can simulate the progress and mechanism of the disease,as well as systemic metabolic disorders such as insulin resistance and oxidative stress.However,it is time-consuming and the fibrosis progression rate is slow.The MCD diet can simulate the serological and pathological features of NASH in 8 weeks,but no obesity or insulin resistance occurred.The CCl_(4)combined with HFD model can ind

关 键 词：非酒精性脂肪肝炎 MCD模型 AMLN模型 胰岛素抵抗 

分 类 号：Q95-33[生物学—动物学]