本文選題：青錢柳多糖　切入點(diǎn)：高脂血癥　出處：《江西農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：為研究青錢柳多糖對(duì)高脂血癥大鼠脂代謝相關(guān)基因DNA甲基化和mRNA表達(dá)的影響,將建模成功的高脂血癥大鼠隨機(jī)分為青錢柳多糖高、中、低劑量組、辛伐他汀組、高脂模型組和空白對(duì)照組,每組分別灌胃青錢柳多糖水溶液400 mg/kg·體重·每天、200 mg/kg·體重·每天、100 mg/kg·體重·每天、4 mg/kg·體重·每天辛伐他汀水溶液,另兩組灌胃等量無(wú)菌水,每組16只,連續(xù)喂養(yǎng)8周。實(shí)驗(yàn)結(jié)束后采用亞硫酸鹽測(cè)序法(Bisulfite Sequencing PCR,BSP)和Q-PCR測(cè)定大鼠肝臟和脂肪組織中瘦素(LEPTIN)、黑素皮質(zhì)素受體-4(MC4R)、微粒體甘油三酯轉(zhuǎn)運(yùn)蛋白(MTTP)、阿片-促黑素細(xì)胞皮質(zhì)素原(POMC)、過(guò)氧化物酶體增殖體激活受體-γ(PPARγ)五個(gè)基因啟動(dòng)子區(qū)域DNA甲基化水平、mRNA表達(dá)情況、甲基轉(zhuǎn)移酶活性以及和它們之間的相關(guān)性,以探究青錢柳多糖在脂代謝方面的分子機(jī)理。1)試驗(yàn)結(jié)果表明,青錢柳多糖(CPP)可以顯著降低高脂血癥大鼠肝臟組織中LEPTIN、MC4R、MTTP、POMC、PPARγ五個(gè)基因啟動(dòng)子區(qū)DNA甲基化水平(P0.05或P0.01);青錢柳多糖可以顯著降低脂肪組織中LEPTIN基因DNA甲基化水平(P0.05),顯著提高POMC基因DNA甲基化水平(P0.05),對(duì)MC4R、MTTP、PPARγ脂肪組織中的DNA甲基化水平無(wú)顯著影響。2)Q-PCR結(jié)果表明,青錢柳多糖(CPP)能夠顯著下調(diào)高脂血癥大鼠肝臟和脂肪組織中LEPTIN、MC4R、MTTP、PPARγ等基因的mRNA表達(dá)量(P0.05或P0.01),對(duì)高脂血癥大鼠肝臟和脂肪中POMC基因mRNA的表達(dá)量無(wú)顯著的影響。3)青錢柳多糖可顯著抑制高脂血癥大鼠脂肪中甲基轉(zhuǎn)移酶(DNMTs)活性(P0.05),對(duì)肝臟中DNMTs活性有一定抑制作用,但效果不明顯;4)相關(guān)性分析結(jié)果顯示,在肝臟組織中,LEPTIN、MC4R、PPARγ、MTTP的DNA甲基化水平與mRNA表達(dá)量無(wú)顯著相關(guān)性,在脂肪組織中,LEPTIN、MC4R、MTTP、POMC的DNA甲基化水平與mRNA表達(dá)量無(wú)顯著相關(guān)性,PPARγ基因呈現(xiàn)極顯著相關(guān),在肝臟和脂肪組織中LEPTIN、MC4R、PPARγ三個(gè)基因DNA甲基化水平與甲基轉(zhuǎn)移酶活性呈顯著正相關(guān),MTTP、POMC基因DNA甲基化水平與甲基轉(zhuǎn)移酶活性正無(wú)顯著相關(guān)性。
[Abstract]:In order to study the effect of Polysaccharide on DNA methylation and mRNA expression of lipid metabolism-related genes in hyperlipidemic rats, the established hyperlipidemic rats were randomly divided into three groups: high, medium, low dose and simvastatin groups. In the hyperlipidemia model group and the blank control group, the rats in each group were administrated with 400 mg/kg / day water of Cyclamen Polysaccharide (400 mg/kg / day) and 100 mg/kg / day (#number0# mg/kg / day) with simvastatin solution (4 mg/kg / day), and the other two groups were given the same amount of aseptic water by stomach. 16 in each group, After 8 weeks of continuous feeding, the rat liver and adipose tissue were determined by bisulfite Sequencing Sequencing BSPP and Q-PCR. The levels of leptin, melanin receptor -4MC-4RN, microsomal triglyceride transporter (TTP), and opioid melanocyte-stimulating cell skin were determined after 8 weeks of continuous feeding. The expression of DNA methylation level in five promoter regions of POMCs, peroxisome proliferator activated receptor-緯 PPAR- 緯, was observed. The activity of methyltransferase and its correlation with them in order to explore the molecular mechanism of lipids metabolism of Polysaccharide of Cyclocylium chinensis showed that, Polysaccharide of Salix Cycariae could significantly reduce the level of DNA methylation in the five promoter regions of LEPTINN MC4RMTTPN POMCPAR緯 in hyperlipidemic rats (P0.05 or P0.01), while Polysaccharide could significantly reduce the level of DNA methylation of LEPTIN gene (P0.05) in adipose tissue of rats with hyperlipidemia. High DNA methylation level of POMC gene (P0.05) had no significant effect on DNA methylation level in MC4RN MTTPPAR- 緯 adipose tissue. Cyclocycline Polysaccharide (CPP) could significantly down-regulate the expression of mRNA in the liver and adipose tissue of rats with hyperlipidemia by P0.01or P0.01P, but had no significant effect on the mRNA expression of POMC gene in liver and fat of hyperlipidemic rats. Glucose could significantly inhibit the activity of methyltransferase (DNMTs) in fat of hyperlipidemic rats and the activity of DNMTs in liver. The results of correlation analysis showed that there was no significant correlation between the DNA methylation level and the expression of mRNA in liver tissue. In adipose tissue, there was no significant correlation between the DNA methylation level and the expression of mRNA. In liver and adipose tissue, there was no significant positive correlation between the DNA methylation level of LEPTINN MC4RPAR- 緯 gene and the activity of methyltransferase. There was no significant positive correlation between DNA methylation level and methyltransferase activity of MTTPPOMC gene in liver and adipose tissue.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R285.5

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