本文關(guān)鍵詞：運(yùn)動及鋅α2糖蛋白干預(yù)對肥胖小鼠肝臟FAS和HSL表達(dá)的影響　出處：《上海體育學(xué)院》2015年碩士論文　論文類型：學(xué)位論文

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【摘要】：【研究目的】:肥胖已成為全球范圍內(nèi)的難題,與糖尿病、心血管疾病、胰島素抵抗癥等疾病有關(guān)。肥胖與脂肪過度堆積密切相關(guān),同時還有其他的因素參與其中,但具體的形成機(jī)制尚未探究清楚。鋅α2糖蛋白(Zinc Alpha2 Glycoprotein,ZAG)是一種脂肪動員因子,它的重要作用之一是促進(jìn)脂肪分解,增加脂肪的利用。本實(shí)驗(yàn)采用了基因表達(dá)的技術(shù),探究運(yùn)動及ZAG干預(yù)對肥胖小鼠肝臟FAS和HSL表達(dá)的影響,探討運(yùn)動及ZAG對肝臟脂代謝影響的機(jī)制,為肥胖防治尋找新的靶點(diǎn),為運(yùn)動及補(bǔ)充外源性ZAG減肥提供科學(xué)依據(jù)�！狙芯糠椒ā�:將pcDNA3.1(-)-mZAG質(zhì)粒轉(zhuǎn)化至TOP10感受態(tài)細(xì)胞中,再用抗藥性標(biāo)記法篩選出含有目的質(zhì)粒的重組體,然后經(jīng)擴(kuò)增、提取、純化獲取大量的ZAG質(zhì)粒,用于轉(zhuǎn)染肥胖小鼠。140只八周齡KM雄性小鼠,隨機(jī)抽取其中的40只小鼠普飼喂養(yǎng),其它小鼠高脂飼料喂養(yǎng),建立肥胖模型。肥胖小鼠隨機(jī)分為4組:安慰劑組(-EP,n=7),ZAG組(-EZ,n=9),運(yùn)動ZAG組(EZ,n=6),運(yùn)動安慰劑組(EP,n=7)。-EP組和EP組注射300ul OPTI-MEM和10ul脂質(zhì)體,-EZ組和EZ組注射300ul OPTI-MEM、10ul脂質(zhì)體和5ug ZAG質(zhì)粒,干預(yù)八周,每周注射兩次;EP組和EZ組,運(yùn)動強(qiáng)度為前四周10m/min、后四周12m/min,5次/周,1h/次。每六天記錄小鼠的體重,實(shí)驗(yàn)第八周末,測定小鼠的體重、脂肪重量。取小鼠肝組織,提取總RNA采用實(shí)時熒光定量PCR法測定小鼠肝臟ZAG、HSL、FAS mRNA表達(dá)水平。用SPSS18.0進(jìn)行統(tǒng)計(jì)分析,所有的數(shù)據(jù)都用平均值±標(biāo)準(zhǔn)差(Mean±S)來表示。兩樣本均值的比較用獨(dú)立樣本T檢驗(yàn),兩個影響因素采用多因素方差分析。雙側(cè)檢驗(yàn)P0.05認(rèn)為有顯著性差異,P0.01認(rèn)為有非常顯著性差異�！狙芯拷Y(jié)果】:1成功擴(kuò)增、提取、純化目的ZAG質(zhì)粒,用于轉(zhuǎn)染肥胖小鼠。2成功構(gòu)建肥胖模型:普通飼料喂養(yǎng)的小鼠體重53.93±2.37g,高脂飼料喂養(yǎng)的小鼠體重67.03±4.68g,高脂小鼠體重是普飼小鼠體重的1.23倍,肥胖模型建立成功。3運(yùn)動和ZAG對小鼠體重、內(nèi)臟脂肪墊、脂體系數(shù)的影響:-EZ組、EZ組小鼠體重下降,-EP組、EP組體重上升,但EP組體重增加量小于-EP組(P0.05);-EZ組、EZ組小鼠體脂及體脂率分別低于-EP組(P0.05)和EP組(P0.05)。4運(yùn)動和ZAG對小鼠體ZAG、HSL、FAS mRNA表達(dá)的影響:-EZ組、EZ組ZAG表達(dá)量分別高于-EP組(P0.05)和EP組(P0.05);EP組、EZ組ZAG表達(dá)量分別高于-EP組(P0.05)和-EZ組(P0.05);-EZ組(P0.05)、EP組(P0.05)HSL表達(dá)量都高于-EP;-EZ組(P0.05)、EP組(P0.05)FAS表達(dá)量都低于-EP組�！狙芯拷Y(jié)論】:1高脂膳食導(dǎo)致小鼠肥胖。2運(yùn)動和ZAG都可降低體重或減緩體重的增加可能是因?yàn)樵黾恿诵∈篌w內(nèi)ZAG的表達(dá),再通過ZAG上調(diào)HSL和下調(diào)FAS的表達(dá)。
[Abstract]:[objective]: obesity has become a global problem, related to diabetes, cardiovascular disease, insulin resistance and other diseases. Obesity and fat accumulation is closely related. At the same time, there are other factors involved, but the formation mechanism is not clear. Zinc 偽 2 glycoprotein zinc Alpha2 Glycoprotein. ZAGG is a kind of fat mobilization factor, one of its important role is to promote fat decomposition, increase the use of fat. To explore the effects of exercise and ZAG intervention on the expression of FAS and HSL in the liver of obese mice, to explore the mechanism of the effects of exercise and ZAG on lipid metabolism of liver, and to find a new target for the prevention and treatment of obesity. To provide a scientific basis for exercise and supplementation of exogenous ZAG to lose weight. [methods]: the pcDNA3.1(-)-mZAG plasmid was transformed into TOP10 receptive cells. A large number of ZAG plasmids were obtained by amplification, extraction and purification. The recombinant plasmid was used for transfection of obese mice into eight weeks old km male mice. The obese mice were randomly divided into 4 groups: placebo group (n = 40), placebo group (n = 40) and ZAG group (n = 4). The rats in the exercise ZAG group and the exercise placebo group were injected with 300ul OPTI-MEM and 10ul liposome respectively. The liposomes of 300ul OPTI-MEMN10ul and 5ug ZAG plasmid were injected into the EZ and EZ groups for eight weeks, twice a week. The exercise intensity of EP group and EZ group was 10m / min for the first four weeks and 12m / min / week for the latter. The body weight of the mice was recorded every six days for 8th weeks. The body weight and fat weight of mice were measured. The total RNA was extracted from the liver tissue of mice and the total RNA was determined by real-time fluorescence quantitative PCR method. The expression level of FAS mRNA was analyzed by SPSS18.0. All the data were represented by mean 鹵standard deviation (mean 鹵S). The mean values of the two samples were compared by independent sample T test. The two influencing factors were analyzed by multivariate analysis of variance. Bilateral test (P0.05) showed significant difference (P0.01). [results] 1: 1 was successfully amplified and extracted. Objective to purify the ZAG plasmid and to construct the obesity model of obese mice by transfection. 2. 2. The weight of mice fed with common feed was 53.93 鹵2.37 g. The body weight of mice fed with high-fat diet was 67.03 鹵4.68g, and the body weight of high-fat mice was 1.23 times that of common feeding mice. The effect of visceral fat pad and lipid system on the weight loss of mice in the EZ group and the weight increase in the EP group, but the weight gain in the EP group was lower than that in the -EP group. The body fat and body fat rate of EZ group were lower than those of -EP group (P0.05), EP group (P0.055.4-exercise) and ZAG. The effect of FAS mRNA expression on ZAG expression in EZ group was higher than that in -EP group (P 0.05) and EP group (P 0.05). The expression of ZAG in EZ group was higher than that in -EP group (P 0.05) and in -EZ group (P 0.05). The expression of HSL in P0.05 + -EZ group was higher than that in -EP5 group. -EZ group (P0.05). EP group (P0.05). The expression of FAS was lower than that of -EP group. [conclusion]:. 1 High fat diet resulted in obesity of both exercise and ZAG could reduce body weight or decrease weight gain, which may be due to the increased expression of ZAG in mice. The expression of HSL and FAS were up-regulated and down-regulated by ZAG.
【學(xué)位授予單位】：上海體育學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：G804.7

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