本文關(guān)鍵詞： miR-92a-3p 3T3-L1前脂肪細(xì)胞 成脂分化 增殖　出處：《華南農(nóng)業(yè)大學(xué)》2016年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：本試驗(yàn)選用3T3-L1前脂肪細(xì)胞系為研究對(duì)象,探討miR-92a-3p對(duì)3T3-L1前脂肪細(xì)胞增殖與分化的影響,旨在探討肥胖癥的發(fā)生機(jī)制,并用化學(xué)修飾模擬物miR-92a-3p mimic和miR-92a-3p inhibitor進(jìn)行轉(zhuǎn)染,運(yùn)用CCK-8檢測(cè)細(xì)胞增殖情況,細(xì)胞流式儀檢測(cè)細(xì)胞周期情況,油紅O染色和甘油三酯檢測(cè),實(shí)時(shí)熒光定量和蛋白免疫印跡法檢測(cè)相關(guān)基因表達(dá)等方法探討miR-92a-3p對(duì)3T3-L1前脂肪細(xì)胞細(xì)胞增殖、細(xì)胞周期、成脂分化及相關(guān)基因和蛋白表達(dá)的變化,得到以下結(jié)果:(1)過(guò)表達(dá)miR-92a-3p抑制3T3-L1前脂肪細(xì)胞增殖,而抑制miR-92a-3p表達(dá)則促進(jìn)3T3-L1前脂肪細(xì)胞增殖。(2)過(guò)表達(dá)miR-92a-3p能通過(guò)下調(diào)細(xì)胞周期標(biāo)志基因(Cyclin D1、Cyclin D3和CDK4)的mRNA和蛋白水平的表達(dá),導(dǎo)致3T3-L1前脂肪細(xì)胞發(fā)生周期G1期阻滯進(jìn)而抑制細(xì)胞增殖,而抑制miR-92a-3p表達(dá)則有相反的作用。(3)過(guò)表達(dá)miR-92a-3p在3T3-L1前脂肪細(xì)胞分化的整個(gè)過(guò)程顯著上調(diào),并呈現(xiàn)時(shí)間依賴(lài)關(guān)系。(4)過(guò)表達(dá)miR-92a-3p,促進(jìn)脂肪細(xì)胞分化標(biāo)志基因(PPARγ、C/EBPα、FABP4)的表達(dá)和甘油三酯的聚積,進(jìn)而促進(jìn)3T3-L1前體脂肪細(xì)胞成脂分化,而抑制miR-92a-3p表達(dá)有相反的效果。
[Abstract]:In this study, 3T3-L1 preadipose cell line was selected as the study object to investigate the effect of miR-92a-3p on the proliferation and differentiation of 3T3-L1 preadipocytes, to explore the mechanism of obesity, and to transfect 3T3-L1 preadipocytes with chemically modified mimics miR-92a-3p mimic and miR-92a-3p inhibitor. Cell proliferation was detected by CCK-8, cell cycle was detected by flow cytometry, oil red O staining and triglyceride were detected. Real-time fluorescence quantitative analysis and Western blotting were used to investigate the effects of miR-92a-3p on the proliferation, cell cycle, adipogenic differentiation and the expression of related genes and proteins in 3T3-L1 preadipocytes. The results showed that the overexpression of miR-92a-3p inhibited the proliferation of 3T3-L1 preadipocytes, while the inhibition of miR-92a-3p expression promoted the proliferation of 3T3-L1 preadipocytes. The overexpression of miR-92a-3p could inhibit the expression of mRNA and protein by down-regulating the cell cycle marker genes: Cyclin D1Cyclin D3 and CDK4). The G 1 phase arrest of 3T3-L1 preadipocytes resulted in the inhibition of cell proliferation, while the inhibition of miR-92a-3p expression had the opposite effect. The overexpression of miR-92a-3p was significantly up-regulated in the whole differentiation process of 3T3-L1 preadipocytes. The expression of miR-92a-3p was overexpressed, which promoted the expression of PPAR- 緯 -C / EBP 偽 -FABP4) and the accumulation of triglyceride, thus promoted the adipocyte adipocyte differentiation of 3T3-L1, but inhibited the expression of miR-92a-3p.
【學(xué)位授予單位】：華南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R589.2

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本文編號(hào)：1503548