本文選題：FADS1 + 高甘油三酯血癥��； 參考：《華中科技大學》2016年碩士論文

【摘要】：目的:通過觀察FADS1在高甘油三酯血癥大鼠肝組織、BRL-3A肝細胞中表達水平及其Cp G島甲基化狀態(tài),探討FADS1在高甘油三酯血癥發(fā)生、發(fā)展中的作用。方法:1、36只雄性SD大鼠,普通飼料適應性喂養(yǎng)1周后隨機分為對照組和高血脂組,分別給予普通飼料及高脂飼料喂養(yǎng),喂養(yǎng)3-5周后測血清中總膽固醇、甘油三酯、低密度脂蛋白和高密度脂蛋白,建立高甘油三酯血癥大鼠模型。再選擇合適濃度的醫(yī)用脂肪乳誘導大鼠肝細胞BRL-3A細胞脂變,檢測細胞中總膽固醇、甘油三酯含量,建立高脂細胞模型,并利用5-Aza-Cd R處理高脂組細胞。2、利用熒光實時定量PCR檢測大鼠肝組織及BRL-3A細胞FADS1 m RNA的表達,酶聯(lián)免疫吸附試驗檢測大鼠血清及BRL-3A細胞上清液中FADS1蛋白的表達情況。3、提取大鼠肝臟組織中DNA后,經(jīng)亞硫酸轉化后行PCR擴增目的基因,并用克隆測序檢測FADS1基因甲基化水平。4、將含有目的基因的質(zhì)粒使用甲基化試劑Sss I處理及未處理后,分別轉染至BRL-3A細胞中,利用熒光素酶報告基因載體系統(tǒng)明確甲基化對FADS1表達的影響。結果:1、高甘油三酯血癥組大鼠血清及高脂BRL-3A細胞中甘油三酯濃度均高于對照組,差異具有統(tǒng)計學意義。2、高甘油三酯血癥組大鼠肝組織及高脂BRL-3A細胞中FADS1 m RNA及蛋白表達均低于對照組,而5-Aza-Cd R處理高脂BRL-3A細胞后,FADS1 m RNA及蛋白表達高于高脂組,差異均有統(tǒng)計學意義。3、FADS1啟動子區(qū)16個甲基化島共6個檢測位點中,高脂組甲基化水平高于對照組,差異均有統(tǒng)計學意義,并且甲基化水平與甘油三酯濃度呈正相關性。4、與甲基化試劑Sss I未處理組相比,處理組熒光酶活性顯著降低,差異具有統(tǒng)計學意義,表明FADS1啟動子區(qū)甲基化抑制了其基因的表達。結論:1、本研究利用高脂飼料喂養(yǎng)雄性SD大鼠,成功建立高甘油三酯血癥動物模型;利用醫(yī)用脂肪乳成功誘導BRL-3A細胞脂變,建立體外模擬高甘油三酯血癥肝細胞模型,且濃度為6%的醫(yī)用脂肪乳為最佳誘導濃度。2、在高甘油三酯血癥大鼠肝組織及脂變BRL-3A細胞中,FADS1基因表達量降低,且甲基化水平高于對照組,FADS1基因啟動子區(qū)甲基化可能參與了高甘油三酯血癥發(fā)病過程。
[Abstract]:Aim: to investigate the role of FADS1 in the pathogenesis and development of hypertriglyceridemia by observing the expression of FADS1 in the liver tissue of hypertriglyceridemia rats and the methylation status of CP G island. Methods Thirty-six male Sprague-Dawley rats were randomly divided into two groups: control group and hyperlipidemia group after one week of adaptive feeding with normal diet. Serum total cholesterol and triglyceride were measured after 3-5 weeks of feeding. The rat model of hypertriglyceridemia was established by low density lipoprotein (LDL) and high density lipoprotein (HDL). Then select the appropriate concentration of medical fat emulsion to induce rat hepatocyte BRL-3A cell lipid change, detect the total cholesterol, triglyceride content in the cells, establish a hyperlipidemic cell model. The expression of FADS1 m RNA in rat liver tissues and BRL-3A cells was detected by real-time fluorescence quantitative PCR. Enzyme linked immunosorbent assay (Elisa) was used to detect the expression of FADS1 protein in rat serum and supernatant of BRL-3A cells. After extracting DNA from rat liver, the target gene was amplified by PCR after sulfite transformation. The methylation level of FADS1 gene was detected by clone sequencing. The plasmid containing the target gene was treated with methylation reagent Sss I and then transfected into BRL-3A cells. Luciferase reporter gene vector system was used to determine the effect of methylation on FADS1 expression. Results the concentration of triglyceride in serum and hyperlipidemic BRL-3A cells in hypertriglyceridemia group was higher than that in control group. The expression of FADS1 m RNA and protein in hypertriglyceridemia group was lower than that in control group, while the expression of FADS1 m RNA and protein in hypertriglyceridemia group was higher than that in hyperlipidemia group. The methylation level in hyperlipidemia group was higher than that in control group, and the difference was statistically significant. There was a positive correlation between methylation level and triglyceride concentration. Compared with the untreated group of methylation reagent Sss I, the activity of fluorescence enzyme in the treatment group was significantly lower than that in the untreated group, and the difference was statistically significant. The results showed that methylation of FADS1 promoter inhibited its gene expression. Conclusion in this study, hypertriglyceridemia animal model was established by feeding male Sprague-Dawley rats with high fat diet, and hypertriglyceridemia hepatocyte model was established by using medical fat milk to induce lipid change of BRL-3A cells. The optimal induction concentration of medical fat milk was 6%, and the expression of FADS1 gene was decreased in hypertriglyceridemia rats' liver tissue and BRL-3A cells, and the expression of FADS1 gene was decreased in hypertriglyceridemia rats, and the expression of FADS1 gene was decreased in hypertriglyceridemia rats. The methylation level of FADS1 gene promoter was higher than that of control group, which may be involved in the pathogenesis of hypertriglyceridemia.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R589.2;R-332

【參考文獻】

相關期刊論文 前10條

1 狄春紅;譚曉華;王小波;吳茵茵;蘭滿;楊磊;;APOA5及FADS1基因多態(tài)性以及單倍體型與血脂水平的關系[J];浙江預防醫(yī)學;2015年11期

2 李藍江;李賀;黃穎;楊瑞;廖澤容;張鴻明;許冰瑩;;GPIHBP1基因rs142861814位點多態(tài)性與高甘油三酯血癥的相關性[J];昆明醫(yī)科大學學報;2015年07期

3 袁云龍;王鋒;汪俊軍;;DNA甲基化與心血管疾病危險因素研究進展[J];臨床檢驗雜志;2014年06期

4 杜憲;焦誼;龍梅;王麗鳳;毛新民;米娜瓦爾·胡加艾;武云;朱曼麗;李琳琳;王燁;;microRNA657與新疆維吾爾族高脂血癥相關性研究[J];新疆醫(yī)科大學學報;2014年06期

5 李銀花;關嘯;魯昆;江龍;王旭;李娜;楊秋;王綠婭;;高三酰甘油血癥合并急性心肌梗死男性患者載脂蛋白A5基因甲基化研究[J];臨床檢驗雜志;2014年01期

6 李研研;王洪云;馬龍樂;韓發(fā)彬;;LPL基因突變與高甘油三酯血癥相關性研究進展[J];中華臨床醫(yī)師雜志(電子版);2014年02期

7 馬蘭;張基昌;崔燕;武軍鐸;崔曉倩;楊金英;尚怡君;;MicroRNAs與冠心病發(fā)生發(fā)展關系的研究進展[J];中國實驗診斷學;2013年07期

8 龐智睿;潘世揚;徐建;王芳;王宏;黃蕾;孫瑞紅;徐娟;韓月;許雨喬;史新惠;;血漿DNA甲基化檢測在宮頸疾病診斷中的應用[J];臨床檢驗雜志;2012年08期

9 王先科;史瑩華;王成章;陳明亮;袁德地;;不同高脂飼料建立高脂血癥大鼠模型的對比研究[J];江蘇農(nóng)業(yè)科學;2012年01期

10 凌宏艷;胡小波;奉水東;楊絲絲;何劍琴;張愷芳;廖端芳;;小鼠間充質(zhì)干細胞脂向分化過程中miRNA-143的表達[J];基礎醫(yī)學與臨床;2012年01期

，

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