目的 探讨高脂饮食(high-fat diet,HFD)对雌性小鼠棕色脂肪蛋白组学的影响.方法 构建高脂饮食诱导的肥胖雌鼠模型.对照组(CON组)小鼠给予普通饮食,实验组(HFD组)小鼠给予高脂饮食,均喂养 21 周,每周记录小鼠体质量,使用小动物体脂分析仪测量身体成分,通过经腹葡萄糖耐量试验(intraperitoneal glucose tolerance test,IPGTT)和胰岛素耐量试验(insulin tolerance test,ITT)评估小鼠葡萄糖耐量和胰岛素耐量.处死小鼠后迅速收集棕色脂肪(brown adipose tissue,BAT)标本于液氮中,后续进行液相色谱-串联质谱检测(liquid chromatogra-phy-tandem mass spectrometry,LC-MS)及蛋白组学数据分析(n=4).结果 与CON组相比,HFD组小鼠体质量增加,体脂含量增加,糖耐量和胰岛素耐量受损.蛋白组学分析以差异倍数(fold change,FC)>1.5 或<0.67,且P<0.05 为标准筛选差异蛋白,聚类分析热图结果显示,HFD小鼠Ucp1 水平升高,蛋白免疫印迹(Western blotting,WB)结果显示,HFD小鼠棕色脂肪中Ucp1 水平更高;亚细胞定位分析表明线粒体是含有差异蛋白数目最多的细胞器;基因本体(gene ontology,GO)功能和京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析显示:与CON组相比,HFD组下调蛋白主要与肌肉收缩有关,涉及钙信号通路等,而上调蛋白主要与脂质分解代谢有关,涉及脂质氧化过程以及产热、氧化磷酸化等通路;蛋白质相互作用分析提示网络中心蛋白主要与脂质合成、脂肪酸β氧化有关.结论 肥胖状态下,雌鼠BAT产热通路激活,并伴有脂肪酸β氧化增强和脂肪合成减弱.
Proteomic analysis of brown adipose tissue in diet-induced obese female mice
Objective To investigate the effects of high-fat diet(HFD)on the proteomics of brown adipose tissue(BAT)in female mice.Methods The obese mice models were established with HFD(HFP group).Mice in the con-trol group were fed with normal diet,while those in the experimental group were fed with HFD.Both groups were fed for 21 weeks and body weight were detected weekly.Body composition was measured with small animal body composi-tion analyzer.The glucose tolerance and insulin tolerance were measured with intraperitoneal glucose tolerance test(IPGTT)and insulin tolerance test(ITT)respectively.After the mice were sacrificed,BAT was quickly collected into liquid nitrogen,and then 4 samples from either group were analyzed with liquid chromatography-tandem mass spectrom-etry(LC-MS)and proteomics.Results Compared with the control group,the HFD group had significantly increased body weight and fat content,but impaired glucose tolerance and insulin tolerance.Differential proteins were screened with FC>1.5 or<0.67 and P<0.05.Cluster heatmap revealed an increased Ucp1 level in HFD mice,and Western blot-ting further confirmed higher Ucp1 expression in HFD mice.Subcellular localization analysis indicated that mitochondria were the organelles containing the largest number of differential proteins.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses showed that the down-regulated proteins in the HFD group were mainly involved with muscle contraction and calcium signaling pathway,while the up-regulated proteins with lipid catabolism,lipid oxidation process,thermogenesis,oxidative phosphorylation and other pathways.Protein interaction analysis suggested that the central proteins of the network were associated with lipid synthesis and fatty acid β oxidation.Conclusion The thermogenic pathway of BAT was enhanced in obese female mice,accompanied with increased oxida-tion of fatty acid β and decreased fat synthesis.
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