高脂饮食性非酒精性脂肪性肝病小鼠肝脏陷窝蛋白-1的变化  

Changes of caveolin-1 in the livers of mice with nonalcoholic fatty liver disease caused by high-fat diet

作　　者：邱艳[1] 陈洪潭[1] 许国强[1] 虞朝晖[1] 滕晓东[1] 姚洪田[1] 

机构地区：[1]浙江大学医学院附属第一医院消化内科,杭州310003

出　　处：《中华消化杂志》2013年第9期616-620,共5页Chinese Journal of Digestion

基　　金：国家自然科学基金（491010-N11026）

摘　　要：目的探索陷窝蛋白-1在高脂饮食性非酒精性脂肪性肝病（NAFLD）发病中的作用。方法给予12只10周龄雄性C57BL／6小鼠高脂高胆固醇饮食14周，建立NAFLD的动物模型作为实验组。同时，普通饮食饲养同种系小鼠6只作为对照组。第14周末处死全部小鼠，比较两组体质量、肝质量和血脂变化。分别用实时荧光定量PCR（qPCR）和Western印迹法检测陷窝蛋白-1在NAFLD小鼠肝组织中的基因和蛋白表达水平变化。苏木精-伊红染色后光学显微镜下观察小鼠肝脏的脂肪变性情况，免疫组织化学染色后观察陷窝蛋白1在肝组织中分布的变化。采用t检验比较两组肝组织中陷窝蛋白-1mRNA及蛋白水平的差异，Mann—whitneyU检验分析不同程度脂肪肝小鼠的肝脏陷窝蛋白-l表达的免疫组织化学积分差异。结果实验组小鼠在高脂高胆固醇饮食14周后全部发生NAFLD，其中重度9只，中度3只。与对照组比较，实验组小鼠的血清TC[（1．940±0．300）mmol／L比（3．771±0．800）mmol／L，t=-3．760]、高密度脂蛋白胆固醇[（1．120±0．066）mmol／L比（2．224±0．420）mmol／L，t=-5．474]、低密度脂蛋白胆固醇[（0．510±0．191）mmol／L比（1．241±0．660）mmol／L，t=3．332]均显著升高（P均〈0．01），但TG差异无统计学意义（P〉0．05）。实验组小鼠肝脏陷窝蛋白-1的mRNA（1．536±0．226比0．980±0．272，t=3．371，P〈0．05）和蛋白水平（0．643±0．240比0．100±0．130，t=-4．847，P〈0．01）均显著高于对照组。免疫组织化学结果显示，表达增加的陷窝蛋白-1主要分布于肝细胞膜、脂滴膜和细胞质中。结论高脂高胆固醇饮食诱导的NAFLD小鼠肝脏陷窝蛋白-1水平上调，可能参与了NAFLD的发病机制。Objective To explore the role of caveolinq in nonalcoholic fatty liver disease （NAFLD） caused by high-fat diet. Methods A total of 12 ten-week-old male C57BL/6 mice were fed with high-fat and high-cholesterol diet for 14 weeks to establish the NAFLD animal model. And six syngeneic mice fed with normal diet at the same time were taken as control. All the mice were sacrificed by the end of 14th week, body weight, liver weight and the changes of serum lipids of the two groups were compared. The changes of caveolin-1 at mRNA and protein levels i.n the liver of mice with NAFLD were detected by quantitative polymerase chain reaction （qPCR） and Western blot. The liver steatosis of the mice was observed under light microscopy after stained by hematoxylin and eosin. The changes of distribution of caveolin-1 in liver were examined by immunohistochemistry. Thedifferences of caveolin-1 at mRNA and protein level in livers between the two groups were compared by t test. The differences of immunohistochemical scores of caveolin-1 expression in the livers of mice with different degree of fatty liver were analyzed by ordinal variables of two independent samples rank sum test analysis. Results After 14 weeks high fat and high-cholesterol diet, all the mice of experiment group developed NAFLD. Nine of which were severe and three were moderate. Compared with the control group, serum total cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol of experiment group significantly increased （（ 1. 940 ± 0. 300） mmol/L vs （3. 771±0. 800） mmol/L, （1. 120±0. 066） mmol/L vs （2. 224±0. 420） mmol/L, （0. 510±0. 191） mmol/L vs （1. 241±0. 660） mmol/L,t -3. 760, -5. 474, -3. 332, all P〈0.01）, however there was no significant difference in triglyceride （P 〉 0. 05）. The caveolin-1 of experiment group significantly increased at mRNA （1. 536±0. 226 vs 0. 980±0. 272, t=3. 371, P〈0.05） and protein levels （0.643±0.240 vs 0. 100±0. 130, t=-4.847, P〈0.01）. T

关 键 词：窖蛋白1 脂肪肝 高脂血症 胆固醇 膳食 小鼠 近交C57BL 

分 类 号：R575.5[医药卫生—消化系统]