一种颠倒进食节律的新的肥胖小鼠模型建立方法  

A novel mouse model of obesity with inverted feeding rhythm

作　　者：陈嘉瑶 李婷 李欣 康栩倩 林冬莹 徐洁华 王桂香[1] 臧林泉[1] CHEN Jiayao;LI Ting;LI Xin;KANG Xuqian;LIN Dongying;XU Jiehua;WANG Guixiang;ZANG Linquan(Guangdong Pharmaceutical University,Guangzhou 510006,China;Dongxuande(Guangzhou)Health Management Technology Co.,Ltd.,Guangzhou 510030)

机构地区：[1]广东药科大学,广东广州510006 [2]东玄德(广州)健康管理科技有限公司,广东广州510030

出　　处：《中国病理生理杂志》2023年第5期951-960,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81873225)。

摘　　要：目的:建立一种成本较低、用时较短的肥胖小鼠模型。方法:雄性昆明小鼠以丙硫氧嘧啶(14.4 mg/kg)生理盐水溶液腹腔注射,颠倒小鼠进食节律,记录摄食饮水量。连续造模9周,以体重、Lee指数、体脂率、皮下脂肪占总脂肪比例和口服葡萄糖耐量实验结果为检测指标;采用酶联免疫吸附测定检测血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平;对小鼠肝脏进行HE染色和油红O染色观察组织形态学及脂滴分布。取小鼠肝脏进行转录组分析以找到该模型的差异表达基因及最显著表达通路。结果:该方法成功使模型组小鼠的体重、Lee指数和体脂率均显著增加(P<0.01),而皮下脂肪占总脂肪比例显著减少(P<0.01)。与对照组相比,模型组小鼠的TG含量显著增加(P<0.01),TC和LDL-C含量显著增加(P<0.05),HDL-C含量显著减少(P<0.05)。HE染色结果显示,模型组小鼠肝脏中央静脉放射状纹理消失,出现核皱缩、细胞肿大、肝细胞内出现脂肪滴并发生气球样病变的情况。油红O染色结果显示,模型组小鼠肝细胞内出现密集的橘红色油滴。KEGG功能富集结果显示,模型组小鼠肝脏内的差异表达基因显著富集在昼夜节律调节通路,显著性排名12,排名第1的是过氧化物酶体增殖物激活受体(PPAR)信号通路。该通路下,差异基因主要富集在以PPARα为核心的脂肪细胞因子信号通路,有7个基因表达上调,11个基因表达下调。结论:该造模方法可能通过抑制PPARα调节脂代谢的相关信号通路,在9周内快速建立肥胖小鼠模型。AIM:To establish an obese mouse model with low cost and short time.METHODS:Male Kun⁃ming mice were injected intraperitoneally with propylthiouracil(14.4 mg/kg)saline solution.Meanwhile,the feeding rhythm was reversed.The amount of food and water intake was recorded.The modeling period was 9 weeks.Body weight,body weight change,Lee index,body fat percentage and subcutaneous fat to total fat ratio were determined,and oral glu⁃cose tolerance test was performed.Moreover,serum levels of triglyceride(TG),total cholesterol(TC),low-density lipo⁃protein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were measured by enzyme-linked immuno⁃sorbent assay.The histomorphological changes and lipid droplet distribution of liver tissues were observed by HE and oil red O staining.The livers of mice were taken for transcriptome analysis,so as to find the differentially expressed genes and the most significantly expressed pathways of the model established by this method.RESULTS:The method success⁃fully resulted in significant increases in body weight,Lee index and body fat percentage(P<0.01),and a significant de⁃crease in subcutaneous fat to total fat ratio in the model mice(P<0.01).Compared with control group,the mice in model group showed a significant increase in serum TG content(P<0.01),significant increases in serum TC and LDL-C levels(P<0.05),and a significant decrease in serum HDL-C content(P<0.05).Liver HE staining results showed central ve⁃nous radial texture loss,nuclear crinkling,cell enlargement,fat droplets within hepatocytes,and ballooning lesions in model group.Hepatic oil red O staining showed dense orange-red oil droplets in the hepatocytes of model group.The KEGG functional enrichment results showed that differentially expressed genes in the livers of model group were signifi⁃cantly enriched in circadian rhythm regulation pathway,ranking the 12th in significance,while peroxisome proliferator-ac⁃tivated receptor(PPAR)signaling pathway was the 1st.Under PPAR signaling pathway,differ

关 键 词：进食节律 肥胖 食源性 小鼠 PPAR信号通路 

分 类 号：R589.2[医药卫生—内分泌] R363.2[医药卫生—内科学]