本文關(guān)鍵詞： 細胞凋亡 棕櫚酸 翻譯調(diào)控腫瘤蛋白(TCTP) 脂肪轉(zhuǎn)移酶CD36 Fyn激酶 肝激酶B1(LKB1) AMP依賴蛋白激酶(AMPK)　出處：《深圳大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:本課題探討在棕櫚酸鈉(PA)的作用下,胰島β細胞中翻譯調(diào)控腫瘤蛋白(TCTP)表達水平的變化以及PA對胰島β細胞增殖和凋亡的影響。結(jié)合高脂喂養(yǎng)的動物實驗,揭示TCTP介導(dǎo)高脂誘發(fā)胰島β細胞凋亡的分子機制以及由PA引發(fā)胰島β細胞凋亡的信號通路。方法:以棕櫚酸鈉(PA)刺激胰島β細胞來模擬體內(nèi)脂肪酸對β細胞的影響;使用高脂飼料喂養(yǎng)C57小鼠,分析小鼠體重和體內(nèi)血糖水平隨高脂飼料喂養(yǎng)時間長短的變化;通過化學(xué)阻斷劑和腺病毒轉(zhuǎn)染目的蛋白基因進入細胞研究相關(guān)蛋白的功能;使用MTT方法檢測細胞的活力;以免疫熒光方法檢測組織及細胞中蛋白的表達、分布以及細胞凋亡;使用Western-Blot檢測不同條件下細胞中蛋白的活性及表達;使用細胞膜蛋白分離方法分析蛋白在細胞膜上的分布情況;使用質(zhì)譜分析確認純化的蛋白。結(jié)果:棕櫚酸鈉(PA)處理INS1E細胞可以增加CD36在細胞膜上的分布,介導(dǎo)細胞對脂肪酸的攝取及信號向細胞內(nèi)傳遞。高脂喂養(yǎng)小鼠以及PA處理細胞可降低胰島β細胞中TCTP的表達。TCTP與胰島β細胞功能密切相關(guān),并介導(dǎo)PA引起的細胞凋亡。PA可通過CD36-Fyn-LKB1-AMPK信號通路下調(diào)TCTP的表達,進而導(dǎo)致細胞凋亡。TCTP的降解可能與其泛素化增加有關(guān)。
[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.
【學(xué)位授予單位】：深圳大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R587.1

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