自由攝食狀態(tài)下不同膳食脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠肥胖相關(guān)指標(biāo)的影響
發(fā)布時(shí)間：2018-07-18 14:27
【摘要】：目的:為多角度全面分析自由攝食狀態(tài)下不同脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)膳食對(duì)肥胖等慢性病相關(guān)危險(xiǎn)因素的影響,本研究通過(guò)用不同脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)飼料對(duì)大鼠進(jìn)行為期10周的喂養(yǎng),期間記錄大鼠的進(jìn)食、體重等一般情況,測(cè)量血壓、血生化指標(biāo),檢測(cè)脂肪因子、食欲調(diào)節(jié)激素水平及其基因表達(dá)情況,了解腸道菌群種類(lèi)及相對(duì)豐度情況,分析各項(xiàng)指標(biāo)在不同組間的差異及其與肥胖發(fā)生的關(guān)系,為進(jìn)一步揭示膳食結(jié)構(gòu)是否與肥胖發(fā)生相關(guān)及科學(xué)指導(dǎo)合理膳食提供基礎(chǔ)參考數(shù)據(jù)。方法:1、自由攝食狀態(tài)下不同膳食脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠血液指標(biāo)及相關(guān)基因表達(dá)的影響70只健康雄性SD大鼠經(jīng)適應(yīng)性喂養(yǎng)3周后,根據(jù)其體重增長(zhǎng)情況隨機(jī)分為7組,每組各10只,分別飼喂低脂高碳水組、低脂高蛋白組、普通組、中脂低碳水組、中脂低蛋白組、高脂低碳水組和高脂低蛋白組飼料共10周。大鼠自由攝食飲水,單籠飼養(yǎng)。每周測(cè)量大鼠體重、體長(zhǎng),正式實(shí)驗(yàn)開(kāi)始第0、5、10周測(cè)量血壓并進(jìn)行內(nèi)眥采血,檢測(cè)血糖、血脂、游離脂肪酸、胰島素、脂聯(lián)素(Adiponectin)、瘦素(Leptin)、脂多糖結(jié)合蛋白(Lipopoysacchride Binding Protein,LBP)、食欲刺激素(Ghrelin)和肥胖抑制素(Obestatin)水平。實(shí)驗(yàn)結(jié)束時(shí)處死大鼠并取腎周脂肪、附睪周脂肪和大網(wǎng)膜脂肪稱(chēng)重,同時(shí)留取附睪周脂肪組織和胃組織,提取各組織總mRNA,反轉(zhuǎn)錄成cDNA進(jìn)行PCR擴(kuò)增,使用實(shí)時(shí)熒光定量PCR方法檢測(cè)脂肪組織中脂聯(lián)素、瘦素基因和胃組織中生長(zhǎng)激素促分泌素受體(Growth Hormone Secretagogue Receptor,GHSR)、G 蛋白偶聯(lián)受體 39(G Protein-coupled Receptor 39,GPR39)基因的表達(dá)情況。2、自由攝食狀態(tài)下不同膳食脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠腸道菌群結(jié)構(gòu)的影響在對(duì)大鼠分組后(第0周),每組隨機(jī)選取6只大鼠,無(wú)菌采集每只大鼠糞便約1.0 g,并在實(shí)驗(yàn)第10周末采用相同方法采集相同大鼠糞便。提取糞便樣本基因組總DNA,檢測(cè)DNA濃度和純度,之后對(duì)樣本DNA進(jìn)行PCR擴(kuò)增、純化。將質(zhì)檢合格的DNA文庫(kù)在Illumina Miseq平臺(tái)上進(jìn)行測(cè)序分析,測(cè)序讀長(zhǎng)為2×300 bp。將所得序列進(jìn)行拼接、質(zhì)控、過(guò)濾,比對(duì)到已知16SrDNA數(shù)據(jù)庫(kù)并進(jìn)行物種分類(lèi)學(xué)分析、α多樣性分析、β多樣性分析和主成分分析等。結(jié)果:1、自由攝食狀態(tài)下不同膳食脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠血液指標(biāo)及相關(guān)基因表達(dá)的影響第10周末,高脂低碳水組和高脂低蛋白組大鼠血清胰島素和胰島素抵抗指數(shù)均高于低脂高碳水組;中脂低蛋白組大鼠血清總膽固醇、低密度脂蛋白膽固醇水平均低于普通組;中脂低蛋白組、高脂低碳水組、高脂低蛋白組大鼠血清高密度脂蛋白膽固醇水平均低于普通組;低脂高碳水組Ghrelin/Obestatin比值低于中脂低蛋白組和高脂低蛋白組。第10周末,低脂高碳水組大鼠脂肪組織脂聯(lián)素mRNA相對(duì)表達(dá)量高于中脂低碳水組,低脂高蛋白組高于普通組和中脂低碳水組;高脂低蛋白組大鼠胃組織GHSR mRNA相對(duì)表達(dá)量高于低脂高蛋白組;普通組大鼠胃組織GPR39 mRNA相對(duì)表達(dá)量低于除低脂高蛋白組的其他組,低脂高蛋白組低于高脂低碳水組。各組大鼠在各時(shí)期平均每日能量攝入量、體重、體長(zhǎng)、Lee's指數(shù)、體脂比、血壓、空腹血糖、甘油三酯、游離脂肪酸、瘦素、脂聯(lián)素、Ghrelin、Obestatin水平以及脂肪組織瘦素基因表達(dá)均無(wú)統(tǒng)計(jì)學(xué)差異。2、自由攝食狀態(tài)下不同膳食脂肪供能比及產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠腸道菌群結(jié)構(gòu)的影響第10周與第0周相比,大鼠腸道菌群出現(xiàn)擬桿菌門(mén)減少、厚壁菌門(mén)增加、變形菌門(mén)增加的趨勢(shì)。第10周,各組之間腸道菌群豐富度和多樣性沒(méi)有差異。中脂低蛋白組大鼠腸道菌群中擬桿菌門(mén)相對(duì)豐度最低,厚壁菌門(mén)與低脂高蛋白組、中脂低碳水組、高脂低碳水組相比顯著升高,擬桿菌門(mén)/厚壁菌門(mén)比值低于低脂高蛋白組和中脂低碳水組;低脂高碳水組大鼠腸道菌群中S24-7相對(duì)豐度高于中脂低蛋白組;低脂高蛋白組Defluviitaleaceae高于高脂低碳水組;中脂低蛋白組VadinBB60高于除高脂低蛋白組外所有組;低脂高碳水組Incertae_Sedis高于中脂低碳水組和高脂低碳水組。結(jié)論:1、不同膳食脂肪供能比和產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)對(duì)大鼠Lee's指數(shù)、體脂比、血壓、血糖、甘油三酯、游離脂肪酸、瘦素、脂聯(lián)素、Ghrelin、Obestatin水平以及脂肪組織瘦素基因表達(dá)等無(wú)影響。2、高脂肪供能比膳食會(huì)引起高密度脂蛋白膽固醇的降低和胰島素抵抗的產(chǎn)生,可能使大鼠處于體重未改變而糖脂代謝紊亂的狀態(tài)。3、高碳水化合物供能比膳食會(huì)提高脂聯(lián)素基因的表達(dá),具體機(jī)制仍需進(jìn)一步研究。4、自由攝食狀態(tài)下,不同脂肪供能比和產(chǎn)能營(yíng)養(yǎng)素結(jié)構(gòu)飼料對(duì)大鼠腸道菌群豐富度和多樣性沒(méi)有影響。5、與高脂肪膳食相比,高碳水化合物供能比膳食更有助于S24-7、D)efluviitaleaceae和incertae_Sedis的增殖,具體生物學(xué)意義有待于進(jìn)一步研究。
[Abstract]:Objective: to analyze the effects of different fat energy supply ratio and productivity nutrient structure diet on obesity and other chronic disease risk factors in the free feeding state. This study was carried out by feeding rats with different fat energy supply ratio and productive nutrient structure feed for 10 weeks, during which the diet and weight of rats were recorded. In the case of blood pressure, blood biochemical index, fat factor, appetite regulating hormone level and gene expression, the species and relative abundance of intestinal flora were understood, the differences among different groups and the relationship with obesity were analyzed, so as to further reveal whether the dietary structure was related to obesity and scientific guidance. The basic reference data of rational diet were provided. Methods: 1, the effects of different dietary fat supply and productivity on the blood indexes and related genes expression in 70 healthy male SD rats were randomly divided into 7 groups after 3 weeks of adaptive feeding, and 10 rats in each group were fed low fat, respectively. High carbon water group, low fat and high protein group, medium fat low carbon water group, low fat low protein group, high fat low carbon water group and high fat low protein group feed for 10 weeks. Rats were free to feed drinking water, single cage feeding. Measure the body weight and body length of rats every week. The blood pressure was measured at the beginning of the 0,5,10 week, and the blood pressure was measured in the inner canthus, blood glucose, blood lipid, free fat were detected. Fatty acid, insulin, adiponectin (Adiponectin), leptin (Leptin), lipopolysaccharide binding protein (Lipopoysacchride Binding Protein, LBP), appetite stimulating hormone (Ghrelin) and obesity inhibin (Obestatin) level. At the end of the experiment, rats were killed and perinenal fat, epididymal fat and omental fat were weighed, and epididymal adipose tissue and epididymal adipose tissue were left at the end of the experiment. Gastric tissue, extract the total mRNA of each tissue, reverse transcripted into cDNA for PCR amplification, and use real-time fluorescence quantitative PCR to detect adiponectin, leptin gene and growth hormone secretocadotropin receptor (Growth Hormone Secretagogue Receptor, GHSR), G protein coupled receptor 39 (G Protein-coupled 39,) gene in the tissues of the stomach The effect of.2, the effect of different dietary fat supply and productivity on the structure of intestinal microflora in rats under free feeding state (zeroth weeks), 6 rats in each group were randomly selected, and the feces of each rat were collected by 1 g, and the feces were collected by the same method at the end of the experiment and the feces were extracted by the same method. Then the total genomic DNA of the sample was collected and the concentration and purity of DNA were detected. Then the sample DNA was amplified and purified by PCR. The qualified DNA library was sequenced and analyzed on the Illumina Miseq platform. The sequence was sequenced by 2 x 300 bp., and the sequence was spliced, the quality control and filtration were compared to the known 16SrDNA database and the taxonomic analysis of the species was carried out. The alpha was much more. Results of sample analysis, beta diversity analysis and principal component analysis. The results were as follows: 1, the effect of different dietary fat donor energy ratio and productivity nutrient structure on blood indexes and related gene expression in rats at free feeding state for tenth weeks, the serum insulin and insulin resistance index of high fat low carbon water group and high fat low protein group were higher than those of low fat. In the carbon and water group, the serum total cholesterol and low density lipoprotein cholesterol levels in the low fat low protein group were lower than those in the normal group, and the high density lipoprotein cholesterol levels in the low fat low protein group, the high fat and low carbon water group and the high fat and low protein group were lower than those in the normal group, and the Ghrelin/Obestatin ratio in the low fat and high carbon water group was lower than the middle fat low protein group and the high fat group. The relative expression of adiponectin mRNA in adipose tissue of low fat and high carbon water group was higher than that of low fat and low carbon water group at the weekend of low fat and high carbon water group, and the low fat and high protein group was higher than that of the normal group and medium fat low carbon water group. The relative expression of GHSR mRNA in the gastric tissue of the high fat and low protein group was higher than that of the low fat and high protein group, and the relative expression of GPR39 mRNA in the gastric tissue of the normal group rats was relative expression. The average daily energy intake, body weight, body length, Lee's index, body fat ratio, blood pressure, fasting blood glucose, triglyceride, free fatty acid, leptin, lipoprotein, Ghrelin, Obestatin, and leptin gene expression in adipose tissue were expressed at all times. There was no statistical difference.2. The effect of different dietary fat supply ratio and productivity structure on intestinal microflora structure of rats under free feeding state tenth weeks compared with zeroth weeks, the intestinal microflora of rats showed the decrease of bacteriobacteria, the increase of the thick wall bacteria gate and the increase of deformable bacteria doors. Tenth weeks, the abundance and diversity of intestinal microflora among the groups There was no difference in sex. The relative abundance of bacilli was the lowest in the intestinal flora of the low fat low protein group, and the thicker wall and the low fat and high protein group, the low carbon water group, the high fat and low carbon water group were significantly higher, the ratio of the bacilli / thick wall bacteria was lower than the low fat high protein group and the medium fat low carbon water group, and the intestinal microflora of the low fat and high carbon water group was S24-7 The relative abundance was higher than that in the low fat and low protein group; the low fat and high protein group Defluviitaleaceae was higher than the high fat and low carbon water group; the VadinBB60 in the low fat and low protein group was higher than all the groups except the high fat and low protein group; the low fat and high carbon water group Incertae_Sedis was higher than the middle fat and low carbon water group and the high fat and low carbon water group. Conclusion: 1, the different dietary fat supply ratio and the productivity nutrients junction are in conclusion. The Lee's index, body fat ratio, blood pressure, blood pressure, blood sugar, triglyceride, free fatty acid, leptin, adiponectin, Ghrelin, Obestatin, and the expression of leptin gene in adipose tissue have no influence on the.2. The high fat supply can cause the decrease of high density lipoprotein cholesterol and the production of insulin resistance, which may make the rats lose weight. The high carbohydrate supply can improve the expression of the adiponectin gene in the state of.3, which can increase the metabolic disorder of glycolipid. The specific mechanism still needs to further study the.4. Under the free feeding state, the abundance and diversity of the intestinal flora of rats have no effect on the abundance and diversity of the intestinal flora, and the high fat diet is higher than the high fat diet, compared with the high fat diet. Carbohydrate supply is more conducive to the proliferation of S24-7, D) efluviitaleaceae and incertae_Sedis than dietary, and the specific biological significance remains to be further studied.
【學(xué)位授予單位】：中國(guó)疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R151
本文編號(hào)：2132187
[Abstract]:Objective: to analyze the effects of different fat energy supply ratio and productivity nutrient structure diet on obesity and other chronic disease risk factors in the free feeding state. This study was carried out by feeding rats with different fat energy supply ratio and productive nutrient structure feed for 10 weeks, during which the diet and weight of rats were recorded. In the case of blood pressure, blood biochemical index, fat factor, appetite regulating hormone level and gene expression, the species and relative abundance of intestinal flora were understood, the differences among different groups and the relationship with obesity were analyzed, so as to further reveal whether the dietary structure was related to obesity and scientific guidance. The basic reference data of rational diet were provided. Methods: 1, the effects of different dietary fat supply and productivity on the blood indexes and related genes expression in 70 healthy male SD rats were randomly divided into 7 groups after 3 weeks of adaptive feeding, and 10 rats in each group were fed low fat, respectively. High carbon water group, low fat and high protein group, medium fat low carbon water group, low fat low protein group, high fat low carbon water group and high fat low protein group feed for 10 weeks. Rats were free to feed drinking water, single cage feeding. Measure the body weight and body length of rats every week. The blood pressure was measured at the beginning of the 0,5,10 week, and the blood pressure was measured in the inner canthus, blood glucose, blood lipid, free fat were detected. Fatty acid, insulin, adiponectin (Adiponectin), leptin (Leptin), lipopolysaccharide binding protein (Lipopoysacchride Binding Protein, LBP), appetite stimulating hormone (Ghrelin) and obesity inhibin (Obestatin) level. At the end of the experiment, rats were killed and perinenal fat, epididymal fat and omental fat were weighed, and epididymal adipose tissue and epididymal adipose tissue were left at the end of the experiment. Gastric tissue, extract the total mRNA of each tissue, reverse transcripted into cDNA for PCR amplification, and use real-time fluorescence quantitative PCR to detect adiponectin, leptin gene and growth hormone secretocadotropin receptor (Growth Hormone Secretagogue Receptor, GHSR), G protein coupled receptor 39 (G Protein-coupled 39,) gene in the tissues of the stomach The effect of.2, the effect of different dietary fat supply and productivity on the structure of intestinal microflora in rats under free feeding state (zeroth weeks), 6 rats in each group were randomly selected, and the feces of each rat were collected by 1 g, and the feces were collected by the same method at the end of the experiment and the feces were extracted by the same method. Then the total genomic DNA of the sample was collected and the concentration and purity of DNA were detected. Then the sample DNA was amplified and purified by PCR. The qualified DNA library was sequenced and analyzed on the Illumina Miseq platform. The sequence was sequenced by 2 x 300 bp., and the sequence was spliced, the quality control and filtration were compared to the known 16SrDNA database and the taxonomic analysis of the species was carried out. The alpha was much more. Results of sample analysis, beta diversity analysis and principal component analysis. The results were as follows: 1, the effect of different dietary fat donor energy ratio and productivity nutrient structure on blood indexes and related gene expression in rats at free feeding state for tenth weeks, the serum insulin and insulin resistance index of high fat low carbon water group and high fat low protein group were higher than those of low fat. In the carbon and water group, the serum total cholesterol and low density lipoprotein cholesterol levels in the low fat low protein group were lower than those in the normal group, and the high density lipoprotein cholesterol levels in the low fat low protein group, the high fat and low carbon water group and the high fat and low protein group were lower than those in the normal group, and the Ghrelin/Obestatin ratio in the low fat and high carbon water group was lower than the middle fat low protein group and the high fat group. The relative expression of adiponectin mRNA in adipose tissue of low fat and high carbon water group was higher than that of low fat and low carbon water group at the weekend of low fat and high carbon water group, and the low fat and high protein group was higher than that of the normal group and medium fat low carbon water group. The relative expression of GHSR mRNA in the gastric tissue of the high fat and low protein group was higher than that of the low fat and high protein group, and the relative expression of GPR39 mRNA in the gastric tissue of the normal group rats was relative expression. The average daily energy intake, body weight, body length, Lee's index, body fat ratio, blood pressure, fasting blood glucose, triglyceride, free fatty acid, leptin, lipoprotein, Ghrelin, Obestatin, and leptin gene expression in adipose tissue were expressed at all times. There was no statistical difference.2. The effect of different dietary fat supply ratio and productivity structure on intestinal microflora structure of rats under free feeding state tenth weeks compared with zeroth weeks, the intestinal microflora of rats showed the decrease of bacteriobacteria, the increase of the thick wall bacteria gate and the increase of deformable bacteria doors. Tenth weeks, the abundance and diversity of intestinal microflora among the groups There was no difference in sex. The relative abundance of bacilli was the lowest in the intestinal flora of the low fat low protein group, and the thicker wall and the low fat and high protein group, the low carbon water group, the high fat and low carbon water group were significantly higher, the ratio of the bacilli / thick wall bacteria was lower than the low fat high protein group and the medium fat low carbon water group, and the intestinal microflora of the low fat and high carbon water group was S24-7 The relative abundance was higher than that in the low fat and low protein group; the low fat and high protein group Defluviitaleaceae was higher than the high fat and low carbon water group; the VadinBB60 in the low fat and low protein group was higher than all the groups except the high fat and low protein group; the low fat and high carbon water group Incertae_Sedis was higher than the middle fat and low carbon water group and the high fat and low carbon water group. Conclusion: 1, the different dietary fat supply ratio and the productivity nutrients junction are in conclusion. The Lee's index, body fat ratio, blood pressure, blood pressure, blood sugar, triglyceride, free fatty acid, leptin, adiponectin, Ghrelin, Obestatin, and the expression of leptin gene in adipose tissue have no influence on the.2. The high fat supply can cause the decrease of high density lipoprotein cholesterol and the production of insulin resistance, which may make the rats lose weight. The high carbohydrate supply can improve the expression of the adiponectin gene in the state of.3, which can increase the metabolic disorder of glycolipid. The specific mechanism still needs to further study the.4. Under the free feeding state, the abundance and diversity of the intestinal flora of rats have no effect on the abundance and diversity of the intestinal flora, and the high fat diet is higher than the high fat diet, compared with the high fat diet. Carbohydrate supply is more conducive to the proliferation of S24-7, D) efluviitaleaceae and incertae_Sedis than dietary, and the specific biological significance remains to be further studied.
【學(xué)位授予單位】：中國(guó)疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R151
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