成熟脂肪细胞中脂联素基因表达的脂肪酸应答

发布时间：2018-06-30 06:07

本文选题：脂肪酸 + 脂联素　；参考：《华中农业大学》2009年硕士论文

【摘要】： 脂肪组织是机体能量代谢和平衡的主要器官,它能合成和存储甘油三脂,并在需要时分解利用脂肪酸来维持机体能量的平衡,脂肪细胞不仅是脂质的储存库,还可分泌许多与糖和脂肪代谢有关的因子,包括瘦素(leptin)、抵抗素(resistin)、脂联素(adiponectin)和肿瘤坏死因子α(TNF-α)等。脂联素是脂肪细胞特异性分泌与胰岛素敏感密切相关的脂肪细胞因子,脂联素与受体结合后具有增强胰岛素敏感性,抗炎,抗糖尿病,抗动脉粥样硬化等生物学效应。脂联素水平的升高对改善疾病状态具有重要的意义,因此深入研究影响其分泌表达的调控因素就显得尤为重要。本研究选用3T3-LI小鼠前脂肪细胞系为体外培养模型,用不同种类、不同浓度游离脂肪酸(FFA)处理体外培养的成熟3T3-L1脂肪细胞,实时荧光定量PCR和Western-blotting分析FFA处理前后脂联素基因mRNA和蛋白质水平的表达差异,以及相关转录因子PPARγ2、SREBP-1c、C/EBPαmRNA表达和TNF-α、IL-6的表达水平,以探讨游离脂肪酸对成熟脂肪细胞脂联素表达的调控作用。 1.分化开始后的3T3-L1细胞从最初的梭形逐渐演变为圆形,细胞体积增大,胞浆内出现由少而多的脂滴,到第10天,平均有90%以上的前体细胞分化为成熟脂肪细胞,高倍镜下可见透亮融合的脂滴,且分化均一,分化效率高。 2.FFAs抑制3T3-L1脂肪细胞脂联素mRNA的表达和分泌,呈时间、剂量依赖性,各种脂肪酸显著影响脂联素蛋白的分泌作用。 3.饱和脂肪酸PA主要通过上调IL-6、TNF-αmRNA的表达抑制脂联素的基因表达和分泌。多不饱和脂肪酸AA、EPA、DHA通过抑制PPARγ2、CREBP-1c的转录活性影响脂联素的水平。 4.各种类型脂肪酸不影响AdipoR1 mRNA的表达;PA、OA处理组分别降低AdipoR2的表达28%、20%,其余各脂肪酸组对AdipoR2无显著影响。
[Abstract]:Adipose tissue is the main organ of energy metabolism and balance. It can synthesize and store triglycerides, and decompose fatty acids when needed to maintain energy balance. It can also secrete many factors related to glucose and fat metabolism including leptin (leptin), resistin (resistin), adiponectin (adiponectin) and tumor necrosis factor- 伪 (TNF- 伪). Adiponectin (adiponectin) is a kind of adipocyte which is secreted by adipocytes which is closely related to insulin sensitivity. Adiponectin binding with receptor has the biological effects of enhancing insulin sensitivity, anti-inflammation, anti-diabetes, anti-atherosclerosis and so on. The increase of adiponectin level plays an important role in improving the state of disease, so it is particularly important to study the regulatory factors that affect the secretion and expression of adiponectin. In this study, 3T3-LI mouse preadipocytes were used as the culture model, and the mature 3T3-L1 adipocytes were treated with different kinds and different concentrations of free fatty acids (FFA). Real-time fluorescent quantitative PCR and Western-blotting were used to analyze the differences in mRNA and protein levels of adiponectin gene before and after FFA treatment, as well as the mRNA expression of PPAR 纬 _ 2nSREBP-1C / EBP 伪 mRNA and the expression level of TNF- 伪 IL-6. To investigate the effect of free fatty acids on adiponectin expression in mature adipocytes. 1. At the beginning of differentiation, 3T3-L1 cells gradually evolved from spindle shape to round shape, the cell volume increased, and the cytoplasm of 3T3-L1 cells changed from less and more lipid droplets. On the 10th day, more than 90% of the precursor cells differentiated into mature adipocytes on average. FFAs inhibited adiponectin mRNA expression and secretion in 3T3-L1 adipocytes in a time-and dose-dependent manner. Fatty acids significantly affected adiponectin secretion. 3. Saturated fatty acid PA inhibits adiponectin gene expression and secretion mainly by up-regulating the expression of IL-6 TNF- 伪 mRNA. Polyunsaturated fatty acid AAPA-DHA affects adiponectin levels by inhibiting the transcriptional activity of PPAR 纬 _ 2 and CREBP-1c. All kinds of fatty acids did not affect the expression of AdipoR1 mRNA.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R341

【参考文献】