运动及锌α2糖蛋白干预对肥胖小鼠肝脏FAS和HSL表达的影响

发布时间：2018-01-03 12:27

本文关键词：运动及锌α2糖蛋白干预对肥胖小鼠肝脏FAS和HSL表达的影响　出处：《上海体育学院》2015年硕士论文　论文类型：学位论文

更多相关文章：运动 锌α2糖蛋白 肥胖肝脏 FAS HSL

【摘要】：【研究目的】:肥胖已成为全球范围内的难题,与糖尿病、心血管疾病、胰岛素抵抗症等疾病有关。肥胖与脂肪过度堆积密切相关,同时还有其他的因素参与其中,但具体的形成机制尚未探究清楚。锌α2糖蛋白(Zinc Alpha2 Glycoprotein,ZAG)是一种脂肪动员因子,它的重要作用之一是促进脂肪分解,增加脂肪的利用。本实验采用了基因表达的技术,探究运动及ZAG干预对肥胖小鼠肝脏FAS和HSL表达的影响,探讨运动及ZAG对肝脏脂代谢影响的机制,为肥胖防治寻找新的靶点,为运动及补充外源性ZAG减肥提供科学依据。【研究方法】:将pcDNA3.1(-)-mZAG质粒转化至TOP10感受态细胞中,再用抗药性标记法筛选出含有目的质粒的重组体,然后经扩增、提取、纯化获取大量的ZAG质粒,用于转染肥胖小鼠。140只八周龄KM雄性小鼠,随机抽取其中的40只小鼠普饲喂养,其它小鼠高脂饲料喂养,建立肥胖模型。肥胖小鼠随机分为4组:安慰剂组(-EP,n=7),ZAG组(-EZ,n=9),运动ZAG组(EZ,n=6),运动安慰剂组(EP,n=7)。-EP组和EP组注射300ul OPTI-MEM和10ul脂质体,-EZ组和EZ组注射300ul OPTI-MEM、10ul脂质体和5ug ZAG质粒,干预八周,每周注射两次;EP组和EZ组,运动强度为前四周10m/min、后四周12m/min,5次/周,1h/次。每六天记录小鼠的体重,实验第八周末,测定小鼠的体重、脂肪重量。取小鼠肝组织,提取总RNA采用实时荧光定量PCR法测定小鼠肝脏ZAG、HSL、FAS mRNA表达水平。用SPSS18.0进行统计分析,所有的数据都用平均值±标准差(Mean±S)来表示。两样本均值的比较用独立样本T检验,两个影响因素采用多因素方差分析。双侧检验P0.05认为有显著性差异,P0.01认为有非常显著性差异。【研究结果】:1成功扩增、提取、纯化目的ZAG质粒,用于转染肥胖小鼠。2成功构建肥胖模型:普通饲料喂养的小鼠体重53.93±2.37g,高脂饲料喂养的小鼠体重67.03±4.68g,高脂小鼠体重是普饲小鼠体重的1.23倍,肥胖模型建立成功。3运动和ZAG对小鼠体重、内脏脂肪垫、脂体系数的影响:-EZ组、EZ组小鼠体重下降,-EP组、EP组体重上升,但EP组体重增加量小于-EP组(P0.05);-EZ组、EZ组小鼠体脂及体脂率分别低于-EP组(P0.05)和EP组(P0.05)。4运动和ZAG对小鼠体ZAG、HSL、FAS mRNA表达的影响:-EZ组、EZ组ZAG表达量分别高于-EP组(P0.05)和EP组(P0.05);EP组、EZ组ZAG表达量分别高于-EP组(P0.05)和-EZ组(P0.05);-EZ组(P0.05)、EP组(P0.05)HSL表达量都高于-EP;-EZ组(P0.05)、EP组(P0.05)FAS表达量都低于-EP组。【研究结论】:1高脂膳食导致小鼠肥胖。2运动和ZAG都可降低体重或减缓体重的增加可能是因为增加了小鼠体内ZAG的表达,再通过ZAG上调HSL和下调FAS的表达。
[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 伪 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 卤standard deviation (mean 卤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 卤2.37 g. The body weight of mice fed with high-fat diet was 67.03 卤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.
【学位授予单位】：上海体育学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：G804.7

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