肥胖时脂肪细胞的丁酰化修饰

发布时间：2018-07-01 15:44

  本文选题：丁酰化 + 乙酰化　； 参考：《新乡医学院》2017年硕士论文

【摘要】：背景近年来,全球肥胖的人数逐年上升,肥胖及其相关的代谢性疾病已成为影响人类健康的主要疾病之一。脂肪细胞数目增加和单个细胞体积增大导致脂肪组织增生。蛋白翻译后修饰(PTMs)引起的分子结构可调节细胞生理活动和疾病进程,是基因表达后生物学研究的主要目标之一,有研究表明PTMs会影响脂肪细胞分化过程中的亚细胞分布,蛋白与蛋白间的相互作用和生物学活性。近期,一种新的PTM,赖氨酸丁酰化修饰在组蛋白中被发现,研究显示丁酰化修饰与精子发育及某些疾病有着密切联系,但至今还没有关于丁酰化修饰与脂肪发育、脂肪细胞分化,肥胖以及代谢疾病的相关报道。目的探讨丁酰化修饰在脂肪细胞分化中的变化,为进一步阐明其作用机制奠定基础。方法1.动物模型的建立:将6周龄C57BL/6品系的小鼠分为两组,一组给予正常饮食,另一组给予高脂肪饮食,分别在高脂肪饮食的1、5、10周处死小鼠取脂肪组织样品。2.体重变化趋势的检测:在小鼠高脂肪饮食的第0周起,每隔一周都要称量体重一次并记录数据。3.Western blot:提取3T3-L1细胞和小鼠脂肪组织(附睾脂肪)蛋白,分别检测丁酰化修饰和乙酰化修饰的表达;4.油红O染色:染在不同分化时期的3T3-L1脂肪细胞,并用显微镜观察其脂滴的表达量。结果1.与正常饮食组相比,高脂饮食使小鼠体重和附睾脂肪组织增加更明显。2.高脂饮食小鼠附睾脂肪组织丁酰化修饰表达随着高脂饮食时间的延长而增强,乙酰化没有这种改变。同时,两组正常饮食小鼠的这两种修饰都没有发现改变;3.油红染色结果显示,加入诱导剂Ⅱ的第6天3T3-L1脂肪细胞始出现脂滴并且随着诱导时间增长脂滴累积增多;4.Western结果显示,蛋白丁酰化修饰和乙酰化修饰在3T3-L1脂肪细胞分化成熟过程中逐渐增强。5.用Forskolin刺激成熟脂肪细胞,与对照组相比,丁酰化修饰和乙酰化修饰随着刺激时间延长而逐渐减弱。结论蛋白丁酰化修饰可能是脂肪细胞分化和脂肪堆积的必要条件。
[Abstract]:Background in recent years, the number of obese people in the world has increased year by year. Obesity and related metabolic diseases have become one of the major diseases affecting human health. An increase in the number of adipocytes and an increase in the volume of individual cells lead to adipose tissue proliferation. The molecular structure induced by protein post-translational modification (PTMs) regulates cellular physiological activity and disease progression, which is one of the main objectives of post-gene expression biology. Some studies have shown that PTMs affect the distribution of subcells in adipocyte differentiation. Protein-protein interaction and biological activity. Recently, a new type of PTM, lysine butyloyl modification, has been found in histone. Studies have shown that butyloyl modification is closely related to sperm development and some diseases, but there is no study on butyloylation modification and fat development and adipocyte differentiation. Obesity and metabolic diseases are reported. Aim to investigate the changes of butyloyl modification in adipocyte differentiation and to lay a foundation for further elucidation of its mechanism. Method 1. Establishment of animal model: C57BL / 6 strain of 6-week-old mice were divided into two groups: one group was given normal diet and the other group was given high-fat diet. Measurement of body weight change trend: from week 0 of high-fat diet, weight was weighed and data was recorded every other week. 3. Western blot: 3T3-L1 cells and mouse adipose tissue (epididymal fat) protein were extracted. The expression of butyloyl modified and acetylated modified was detected respectively. Oil red O staining: 3T3-L1 adipocytes were stained at different differentiation stages and their lipid droplets were observed by microscope. Result 1. Compared with normal diet group, high fat diet increased body weight and epididymal adipose tissue more significantly. 2. 2. The modified expression of butyloy in epididymal adipose tissue of high fat diet mice increased with the prolongation of high fat diet time, but acetylation did not. At the same time, the two groups of normal diet mice did not find any changes in the two modifications. The results of oil red staining showed that lipid droplets first appeared in 3T3-L1 adipocytes on the 6th day after the addition of inducer 鈪，

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