基于代谢组学及基因芯片研究褪黑素对大鼠生长及代谢的影响

发布时间：2018-01-26 00:00

本文关键词： 褪黑素大鼠代谢组学基因芯片　出处：《河南农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：为研究褪黑素(melatonin,MT)对动物生长及代谢的影响。本试验选择日龄、体重接近的SD级雄性大鼠50只,随机平均分为5组,每组10只,分别为正常日粮组(ND)、正常日粮+褪黑素组(ND+MT)、脂肪添加组(FS)、脂肪添加+低剂量褪黑素组(FS+LMT)和、脂肪添加+高剂量褪黑素组(FS+HMT)。FS组在正常饲粮中添加10%的猪油,ND+MT、FS+LMT、FS+HMT组除饲喂特定饲粮外,分别灌服10mg/kgBW、2mg/kgBW、20mg/kg BW的MT溶液。试验过程中每天称重,计算日增重。试验结束后采血,检测大鼠血液中相关生化指标、激素等含量。剖取内脏主要组织器官并称量相关脂肪组织重量,此外,取肝脏组织做冰冻切片,油红O染色,观察脂肪含量。对ND、ND+MT、FS、FS+MT组大鼠肝脏样品采用1H-NMR的代谢组学方法进行代谢组分析,从小分子代谢物层面反映肝脏营养代谢状态。该技术可鉴定生物体在不同生理状态下的生物标志物,有利于判断生物体在各种环境条件下的代谢状态。在MT作用下,动物机体各器官,包括肝脏的各种代谢途径均可受到影响,相关代谢产物必然发生不同程度的变化,用代谢组学的方法可以全面准确地检测到MT影响肝脏代谢的主要标志物,判断其影响肝脏代谢的途径和程度。ND、ND+MT组大鼠肝脏样品采用基因芯片的方法,从基因层面进一步探究MT对大鼠的影响。本研究可以得出如下结果:(1)ND+MT与ND相比日均采食量、平均日增重、肝脏重显著下降(P0.05);肝脏组织主要脂肪酸、总胆固醇、大网膜含量、肾周脂肪、附睾脂肪含量均显著降低(P0.05)。FS+LMT组日均采食量、平均日增重、肝脏主要脂肪酸、大网膜含量、附睾脂肪含量较FS组降低,肾周脂肪含量显著升高(P0.05);FS+HMT组的大网膜重、附睾脂肪含量较FS组显著降低(P0.05)。FS组附睾脂肪含量较ND组显著降低(P0.05),而肾周脂肪含量则较ND组显著增加(P0.05);FS+LMT与FS+HMT相比表明,高剂量褪黑素处理组腹腔内脂肪的含量升高。(2)通过核磁共振的方法进行代谢组学研究,可检测到的代谢产物共有63种,分别为氨基酸类,有机酸类,糖类,多胺类以及维生素类,其中共发现13中潜在标志性代谢物,其中ND组与ND+MT组之间共有5种,FS与FS+LMT组共有8种,ND与FS组共有5种,这些代谢物主要与氨基酸、脂肪和碳水化合物等代谢途径有关,可作为褪黑素对大鼠肝脏影响的标志物。(3)通过ND组与ND+MT组大鼠肝脏的基因芯片分析,结合代谢组学的试验结果,发现与代谢组学结果相关的差异性表达的基因共有16个,分别属于糖代谢,脂肪酸代谢,氨基酸代谢的通路,这也从基因的角度与代谢组学构成了互相印证,使得两种试验方法互相补充,也更加科学全面的研究了褪黑素对大鼠代谢的影响。本研究可以得出如下结论:(1)相同的日粮条件下MT可以降低大鼠的生长速度,减少大鼠肝脏内脂肪的含量,改变脂肪在体内的分配;(2)采用1H-NMR的代谢组学方法可以宏观地观察到肝脏在MT的作用下的代谢状态的变化,为科学全面的阐述肝脏在受到褪黑素影响下的代谢情况提供依据;(3)MT可以加强大鼠肝脏物质代谢通路中的关键酶基因的表达。
[Abstract]:To study the influence of melatonin (melatonin, MT) on the growth and metabolism of the animal. This study selected age, weight close male SD 50 rats were randomly divided into 5 groups, 10 rats in each group were normal diet group (ND), normal diet + melatonin group (ND+MT). Add fat group (FS), add fat + low dose of melatonin group (FS+LMT) and fat, add + high dose of melatonin group (FS+HMT).FS was added 10% lard in normal diet, ND+MT, FS+LMT, FS+HMT group were fed diet specific, was administered 10mg/kgBW, 2mg/kgBW, MT 20mg /kg BW solution. During the test day weighed to calculate daily gain. At the end of the experiment were detected in rat blood biochemical index, hormone profile. Main organs and weighing related visceral adipose tissue weight, in addition, liver tissue slices, oil red O staining observation. The fat content of ND. ND+MT, FS, FS+ The metabolism of group 1H-NMR metabolic group analysis method liver samples of rats in group MT with small molecular metabolites levels reflect the liver nutritional and metabolic status. This technique can identify organisms in different physiological conditions of biomarkers in various environmental conditions conducive to the metabolism of organisms. State estimation in the presence of MT in different organs of animal the body, including various metabolic pathways of the liver can be affected by the metabolites must change to different degree, using the method of metabonomics can comprehensively and accurately detect MT main sign of liver metabolism, determine the effect of liver metabolism pathway and.ND method, the liver samples of rats in ND+MT group by gene the chip, further explore the effects of MT on rats from the gene level. We can draw the following results: (1) ND+MT ND compared with the average daily feed intake, average daily weight gain, liver Weight decreased significantly (P0.05); the main fatty acids in liver tissue, total cholesterol content, omentum, perirenal fat, epididymal fat content decreased significantly (P0.05) in group.FS+LMT, the average daily feed intake, average daily gain, liver fatty acid content, omentum, epididymal fat content was lower than that of FS group, perirenal fat the content was significantly increased (P0.05); FS+HMT group of omental weight, epididymal fat content decreased significantly compared with group FS (P0.05).FS group, epididymal fat content decreased significantly compared with ND group (P0.05), and perirenal fat content was significantly higher than that in ND group (P0.05); FS+LMT compared with FS+HMT showed that increased the content of high dose melatonin treatment group intraperitoneal fat. (2) studied by NMR method for metabolic metabolites could be detected in a total of 63, respectively, amino acids, organic acids, sugars, vitamins and polyamines, which were found in the metabolic potential marker 13 Among them, between ND group and ND+MT group were 5, FS and FS+LMT were 8, ND and FS group, a total of 5 species, mainly related to these metabolites and amino acids, fat and carbohydrate metabolism, markers can be used as the effect of melatonin on rat liver by gene chip (3). ND group and ND+MT group rat liver test results analysis, combination of metabolomics, discovery and metabolomics results related genes were 16, belonging to the sugar metabolism, fatty acid metabolism, amino acid metabolism pathway, which is from the perspective of gene and generation of metabolomics constitute each other that makes two kinds of test methods complement each other, but also more scientific and comprehensive study of the effects of melatonin on the metabolism of rats. The conclusions were as follows: (1) the same diet under the condition of MT can reduce the growth rate of rats, reduce the content of fat in the liver of rats, Change in the body fat distribution; (2) using 1H-NMR metabonomic method of macroscopic observed changes in the metabolic state under the effect of MT in the liver, provide a scientific basis for the comprehensive exposition of metabolism in liver by melatonin under the influence; (3) MT can enhance the expression of key enzyme genes in the liver metabolism pathway in rats.

【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.2

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