多不饱和脂肪酸对3T3-L1脂肪细胞白色脂肪棕色化的作用及机制研究

发布时间：2018-08-08 14:40

【摘要】：研究目的肥胖及其相关代谢紊乱疾病问题日益突出,目前缺少行之有效的减肥方法。人体内的棕色脂肪组织因为可以促进能量代谢,调节葡萄糖和脂质水平,成为研究热点。n-3和n-6多不饱和脂肪酸(PUFAs)能够调节脂肪细胞分化和功能,改善肥胖。本研究旨在采用n-3系的二十二碳六烯酸(Docosahexaenoicacid,DHA)、二十碳五烯酸(Eicosapentaenoic Acid,EPA),以及 n-6 系的花生四烯酸(Arachidonic acid,AA)处理各个阶段的3T3-L1脂肪细胞,观察其糖脂水平与棕色化相关标志基因的表达,以及线粒体数量变化,探讨n-3、n-6两类多不饱和脂肪酸对白色脂肪的棕色化作用及机制。研究方法采用浓度为10 μ和100 μM的DHA、EPA、AA分别处理3T3-L1各阶段的脂肪细胞。用试剂盒检测葡萄糖消耗量与甘油三酯水平,用Q-PCR和蛋白免疫印迹法检测棕色化标记基因的mRNA与蛋白表达量,,Q-PCR和Mito-Tracker Green染色法检测线粒体含量。结果DHA处理3T3-L1前脂肪细胞2天后,出现了白色脂肪棕色化的细胞表征。DHA和EPA处理处于分化阶段的3T3-L1脂肪细胞8天后,棕色脂肪标志性基因UCP1蛋白表达量显著增加,线粒体含量和葡萄糖消耗量增加。100 μ作用浓度下,DHA处理组的3T3-L1脂肪细胞甘油三酯水平下降,EPA处理组的细胞甘油三酯水平升高。EPA和AA处理已经完全分化成熟的脂肪细胞2天后,出现了棕色脂肪细胞表征,但是线粒体数目没有变化。在3T3-L1细胞诱导分化过程中,DHA在分化早期促进白色脂肪细胞棕色化,而EPA在分化后期促进白色脂肪细胞棕色化。结论DHA、EPA和AA在10μM和1000μM浓度下均可以促进不同阶段的3T3-L1脂肪细胞获得棕色脂肪细胞的表型,促进其线粒体生成以及葡萄糖消耗。在合适时机补充膳食PUFAs可以促进白色脂肪棕色化,可作为肥胖预防和控制的一个潜在靶标。
[Abstract]:Objective the problem of obesity and related metabolic disorders is becoming more and more prominent, and there is a lack of effective weight loss methods. Brown adipose tissue can promote energy metabolism, regulate glucose and lipid levels, and become a research hotspot. N-3 and n-6 polyunsaturated fatty acid (PUFAs) can regulate adipocyte differentiation and function, and improve obesity. The aim of this study was to observe the expression of glycolipid level and browning related marker genes in 3T3-L1 adipocytes treated with n-3 lines of 22 carbohexaenoic acid, Eicosapentaenoic acidaenoic acid and n-6 arachidonic acid. And the change of mitochondria number, the browning effect and mechanism of n-3 n-6 polyunsaturated fatty acids on white fat were discussed. Methods the adipocytes of 3T3-L1 were treated with 10 渭 M and 100 渭 M DHA EPAAA, respectively. Glucose consumption and triglyceride levels were measured by kit, mRNA and protein expression of brown labeled genes were detected by Q-PCR and Western blotting. Mitochondrial contents were detected by Q-PCR and Mito-Tracker Green staining. Results after 2 days of 3T3-L1 treatment with DHA, the brown cells with white fat were browned. DHA and 3T3-L1 adipocytes in differentiation stage were treated with EPA for 8 days. The expression of UCP1 protein of brown fat marker gene was significantly increased. The decrease of triglyceride level in 3T3-L1 adipocytes in DHA treatment group. The level of triglyceride in 3T3-L1 adipocytes was increased. Brown fat cells were present, but the number of mitochondria did not change. During the differentiation of 3T3-L1 cells, DHA promoted the browning of white adipocytes at the early stage of differentiation, while EPA promoted the browning of white adipocytes at the later stage of differentiation. Conclusion at 10 渭 M and 1000 渭 M, both DHAA and AA can promote the formation of brown adipocytes, mitochondrial formation and glucose consumption in 3T3-L1 adipocytes at different stages. Dietary PUFAs supplementation at the appropriate time can promote the browning of white fat and can be used as a potential target for the prevention and control of obesity.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.2

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