TCTP介导高脂诱发胰岛β细胞凋亡的分子机制研究

发布时间：2018-02-11 11:22

本文关键词： 细胞凋亡棕榈酸翻译调控肿瘤蛋白(TCTP) 脂肪转移酶CD36 Fyn激酶肝激酶B1(LKB1) AMP依赖蛋白激酶(AMPK)　出处：《深圳大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本课题探讨在棕榈酸钠(PA)的作用下,胰岛β细胞中翻译调控肿瘤蛋白(TCTP)表达水平的变化以及PA对胰岛β细胞增殖和凋亡的影响。结合高脂喂养的动物实验,揭示TCTP介导高脂诱发胰岛β细胞凋亡的分子机制以及由PA引发胰岛β细胞凋亡的信号通路。方法:以棕榈酸钠(PA)刺激胰岛β细胞来模拟体内脂肪酸对β细胞的影响;使用高脂饲料喂养C57小鼠,分析小鼠体重和体内血糖水平随高脂饲料喂养时间长短的变化;通过化学阻断剂和腺病毒转染目的蛋白基因进入细胞研究相关蛋白的功能;使用MTT方法检测细胞的活力;以免疫荧光方法检测组织及细胞中蛋白的表达、分布以及细胞凋亡;使用Western-Blot检测不同条件下细胞中蛋白的活性及表达;使用细胞膜蛋白分离方法分析蛋白在细胞膜上的分布情况;使用质谱分析确认纯化的蛋白。结果:棕榈酸钠(PA)处理INS1E细胞可以增加CD36在细胞膜上的分布,介导细胞对脂肪酸的摄取及信号向细胞内传递。高脂喂养小鼠以及PA处理细胞可降低胰岛β细胞中TCTP的表达。TCTP与胰岛β细胞功能密切相关,并介导PA引起的细胞凋亡。PA可通过CD36-Fyn-LKB1-AMPK信号通路下调TCTP的表达,进而导致细胞凋亡。TCTP的降解可能与其泛素化增加有关。
[Abstract]:Aim: to investigate the changes of TCTP expression in islet 尾 cells induced by sodium palmitate (PAA) and the effect of PA on proliferation and apoptosis of islet 尾 cells. To reveal the molecular mechanism of TCTP mediated apoptosis of islet 尾 cells induced by hyperlipidemia and the signal pathway of PA inducing apoptosis of islet 尾 cells. Methods: sodium palmitate was used to stimulate 尾 cells to simulate the effect of fatty acids on 尾 cells in vivo. C57 mice were fed with high fat diet to analyze the changes of body weight and blood glucose level with the feeding time of high fat diet, and to study the function of related proteins by transfection of target protein gene with chemical blocker and adenovirus. MTT method was used to detect cell viability, immunofluorescence method was used to detect the expression, distribution and apoptosis of proteins in tissues and cells, and Western-Blot was used to detect the activity and expression of proteins in cells under different conditions. Cell membrane protein separation was used to analyze the distribution of the protein on the cell membrane, and the purified protein was confirmed by mass spectrometry. Results: INS1E cells were treated with sodium palmitate to increase the distribution of CD36 on the cell membrane. High fat feeding mice and PA treated cells can reduce the expression of TCTP in islet 尾 cells. TCTP is closely related to the function of islet 尾 cells. PA-induced apoptosis. PA can down-regulate the expression of TCTP through CD36-Fyn-LKB1-AMPK signaling pathway, which may be related to the increase of Ubiquification.
【学位授予单位】：深圳大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.1

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