有氧运动对肥胖小鼠棕色脂肪mRNA表达谱的影响
Effect of Aerobic Exercise on Gene Expression Profile of Brown Adipose Tissue in Obese Mice

摘要：目的：通过mRNA表达谱芯片筛选有氧运动前后肥胖小鼠棕色脂肪差异表达基因及通路，探究运动活化棕色脂肪机制。方法：30只雄性C57BL/6J小鼠分为：普通对照组（N，n=8），肥胖对照组（OB，n≥6）和肥胖运动组（E，n≥6）。E组进行10 m/min，1 h/d，6 d/周的跑台训练。每周记录各组摄食量和体重。4周后，测试血糖血脂，取肩胛处棕色脂肪，提取的RNA逆转录合成cDNA后进行mRNA表达谱芯片杂交扫描和生物信息学分析。采用倍数法（≥2，P≤0.05）筛选上下调差异表达基因，用KOBAS2.0软件对差异基因进行基因功能注释和信号通路富集度统计分析。筛选参与关键生物过程和信号通路的重要差异基因进行RT-PCR验证。结果：与N组相比，OB组棕色脂肪组织上调的基因有445个，下调基因有796个，差异基因的生物功能主要集中在脂质、固醇，胆固醇和类固醇的合成代谢过程及白细胞和粒细胞趋药性过程；与OB组相比，E组的上调基因有486个，下调基因有286个，差异基因的生物过程主要集中在脂质，脂肪酸，固醇和胆固醇的合成代谢过程、羧酸和酮酸的代谢过程、辅酶的代谢过程和抗氧化过程；运动调节棕色脂肪功能的通路主要集中在PPAR，AMPK，Insulin和VEGF信号通路等过程。结论：有氧运动可能通过调节一系列基因和通路来提高棕色脂肪活性，促进肥胖机体减肥。
Abstract: Objective: The purposes of this study were to explore the differentially expressed genes and pathways of brown adipose tissue in obese mice before and after aerobic exercise by gene expression profile, and to find the mechanism of exercise activating BAT. Methods: Thirty male C57BL/6J mice were divided into control group (N, n=8), high-fat diet control group (OB, n≥6) and high-fat diet exercise group (E, n≥6). Group E participated in treadmill training (10m/min, 1h/d, 6d/w). The food intake and body weight of all the mice were recorded weekly. After 4 weeks, blood lipid and blood glucose were detected; RNA was extracted from BAT of scapula, cDNA was synthesized, and microarray hybridization were scanned to generate gene lists with corresponding fluorescence intensity data. Genes that were significantly (P≤0.05) up- or down- regulated more than 2 folds were chosen to be analyzed based on gene ontology and enrichment by KOBAS2.0, and the targeted genes that played in key biological process (BP) and pathway were selected and tested by RT-PCR. Results: Comparing with group N, there are 445 genes significantly up regulated and 796 down regulated in group OB. The BP of these genes are mainly about the synthesis and metabolism process of lipids, sterol, cholesterol and steroid, as well as the chemotaxis of hemameba and granulocyte. Comparing with group OB, there are 486 genes significantly up regulated and 286 down regulated. The BP of these genes are mainly about the synthesis and metabolism process of lipids, sterol, cholesterol and steroid, as well as the metabolism process of carboxylic acid, acetone acid and coenzyme, and antioxidant process. The pathways that aerobic exercise regulates the function of BAT mainly are PPAR, AMPK, Insulin and VEGF signaling pathways. Conclusion: Aerobic exercise may improve the activity of BAT by regulating genes and pathway, to promote weight loss of obese mice