间歇性断食对肠道菌群及宿主代谢的影响

发布时间：2018-06-27 18:38

本文选题：断食 + 肠道菌群　；参考：《南方医科大学》2017年硕士论文

【摘要】：目的:1.研究间歇性断食对老年前期肥胖大鼠代谢及肠道菌群的影响。2.研究不同断食形式对人体肠道菌群的影响。3.研究饮水断食对人体代谢的影响。方法:1.雌性Wistar大鼠经42w高脂高糖饲料饲养造模,根据体质量(x±SD)选取模型鼠进行间歇性断食干预。干预方法为每2w断食72h,总干预时间18w。干预后进行口服葡萄糖耐量试验、血脂四项检测。收集粪便,通过Illumina高通量测序检测16S rRNA基因V4可变区,运用BIPES、QIIME及LEfSe分析肠道菌群。2.招募健康志愿者进行持续7日的断食实验,分为饮水断食组和果汁断食组。收集断食前两周至断食后四周的粪便样本,通过Illumina高通量测序检测16S rRNA基因V4可变区,运用BIPES、QIIME及LEfSe分析肠道菌群。3.收集饮水断食组志愿者在断食前后的血液标本,进行生理生化指标检测以及预处理后进行核磁共振分析样本中的代谢产物。使用MestReNova对1H CPMG谱进行分析,Simca-P进行多元统计分析。根据人类代谢产物数据库对断食前后有统计差异的代谢产物成分进行鉴定。结果:1.大鼠实验中间歇性断食组体质量相对于模型对照组显著下降(P0.01);高密度脂蛋白胆固醇和低密度脂蛋白胆固醇均显著下降(P0.05);空腹血糖显著上升(P0.01);葡萄糖耐量测试曲线下面积显著高于模型对照组,糖耐量减退(P0.05);HE染色显示间歇性断食轻度减少肝脏脂肪变性。肠道菌群结果显示,断食组肠道菌群得到显著改善,具体表现为YS2、RF32、Helicobacteraceae(螺杆菌科)增加,Lactobacillus(乳杆菌属)、Roseburia(罗氏菌属)、Erysipelotrichaceae(韦荣球菌科)、Ralstonia(青枯菌属)、Bradyrhizobiaceae(慢生根瘤菌科)和RF39减少。Spearman相关性分析发现Bradyrhizobiaceae与总胆固醇、高密度脂蛋白胆固醇呈正相关;RF39与大鼠体质量呈正相关。2.饮水断食比果汁断食对人肠道菌群的影响更为强烈,且效果在宿主恢复饮食后能长久维持。但饮水断食的效果具有个体差异,总的来看饮水断食能使肠道菌群变成以Prevotella(普雷沃氏菌属)、Bacteroides(拟杆菌属)、Faecalibacterium(柔嫩菌属)和Succinivibrio(琥拍酸弧菌属)为主。3.饮水断食前后的代谢产物水平存在明显差异,断食期间3-羟基丁酸酯和乙酰乙酸的表达上升(P0.05),而丙氨酸、谷氨酸盐、组氨酸、氧化三甲胺、β-葡萄糖、α-葡萄糖的表达显著下降(P0.05)。断食期间,机体的血糖、胰岛素、甘油三酯下降,酮体、总胆固醇、低密度脂蛋白胆固醇、血肌酐、血尿酸、皮质醇升高,提示机体可能通过促进代谢脂肪酸、肌肉、核酸等物质产生能量。机体代谢改变可能与脂肪酸合成、鸟氨酸循环、氨基酸代谢、丙氨酸循环、转录翻译、酮体代谢、三羧酸循环、糖异生、糖酵解等多个通路相关。在恢复饮食3天后代谢产物水平恢复至与断食前相似。结论:1.间歇性断食能改善肠道菌群,降低老年前期肥胖大鼠体质量和血脂水平,但对糖代谢可能有不良影响。2.无卡路里摄入比限制卡路里摄入的断食方式对肠道菌群影响更大。3.饮水断食能把拟杆菌为主的肠道菌群变为以普雷沃氏菌为主,而对普雷沃氏菌为主的肠道菌群结构影响不大。4.机体在饮水断食期间可能通过调节能量相关的代谢通路,消耗脂肪酸、肌肉、核酸维持机体活动所需的能量消耗。5.饮水断食后恢复饮食第3天,机体的代谢产物都恢复到正常水平,但肠道菌群仍没有恢复,说明机体代谢调节比肠道菌群对饮食的改变更加敏感。总体而言,断食对肠道菌群的影响个体差异性较为显著,用于临床疾病预防和治疗仍需深入研究,但能为肥胖患者提供新的视角。
[Abstract]:Objective: 1. study the effect of intermittent feeding on metabolism and intestinal microflora in obese rats.2. study the influence of different forms of food break on human intestinal flora.3. study on the effect of drinking water break on human metabolism. Methods: 1. female Wistar rats were fed with 42W high fat and high sugar diet, and the model rats were selected according to body mass (x + SD). Intermittent fasting intervention. The intervention method was 72h per 2W, oral glucose tolerance test and four blood lipid tests. Feces were collected, V4 variable region of 16S rRNA gene was detected by Illumina sequencing, and BIPES, QIIME and LEfSe analysis of intestinal flora.2. were used for 7 day continuous feeding. The experiment was divided into the drinking water feeding group and the fruit juice group. The faecal samples were collected from two weeks to the four weeks after the disconnection. The 16S rRNA gene V4 variable region was detected by Illumina high throughput sequencing, and BIPES, QIIME and LEfSe were used to analyze the intestinal microflora.3. to collect the blood specimens before and after the broken food in the drinking water group, and the physiological and biochemical indexes were examined. The metabolites in the nuclear magnetic resonance analysis samples were measured and pretreated. The 1H CPMG spectrum was analyzed with MestReNova, and the Simca-P was analyzed by multivariate statistical analysis. According to the human metabolite database, the metabolites of the statistical differences before and after the disconnection were identified. Results: the mass of the intermittent feeding group in the 1. rats was relative. In model control group (P0.01), high density lipoprotein cholesterol (HDL) and low density lipoprotein cholesterol (HDL) were significantly decreased (P0.05), fasting blood glucose increased significantly (P0.01), and the area under glucose tolerance test curve was significantly higher than that of model control group, impaired glucose tolerance (P0.05), and HE staining showed intermittent intermittent feeding reduced liver steatosis slightly. The intestinal flora showed that the intestinal microflora of the diet group improved significantly, including YS2, RF32, Helicobacteraceae (helicobacteraceae), Lactobacillus (Lactobacillus), Roseburia (Roche), Erysipelotrichaceae (Wei Rong family), Ralstonia (Rhizoctonia), Bradyrhizobiaceae (slow Rhizobiaceae) and RF39 reduced.Spearman Correlation analysis found that Bradyrhizobiaceae was positively correlated with total cholesterol and high density lipoprotein cholesterol; RF39 was positively related to the body mass of rats;.2. was more strongly influenced by the drinking water break than the fruit juice feeding on the human intestinal flora, and the effect could be maintained for a long time after the host was restored to the diet. Drinking water break can make intestinal flora become Prevotella (Poulet Was bacteria), Bacteroides (bacteriobacterium), Faecalibacterium (tenella) and Succinivibrio (Vibrio succinate), the metabolic products of.3. before and after drinking water are significantly different, and the expression of 3- hydroxybutyrate and acetoacetic acid increases during the period of feeding (P0.05). The expression of alanine, glutamate, histidine, trimethylamine, beta glucose, and alpha glucose decreased significantly (P0.05). During the diet, the body's blood sugar, insulin, triglyceride, ketone body, total cholesterol, low density lipoprotein cholesterol, serum creatinine, blood uric acid, cortisol increased, suggesting that the body may promote the metabolism of fatty acids and muscles. Meat, nucleic acid and other substances produce energy. Metabolic changes may be related to fatty acid synthesis, ornithine cycle, amino acid metabolism, alanine cycle, transcription translation, ketone metabolism, three carboxylic acid cycle, sugar isogenesis, glycolysis, etc. after 3 days of recovery, the level of metabolite recovery is similar to that before eating. Conclusion: 1. intermittent feeding It can improve the intestinal flora and reduce the body mass and blood lipid level of the obese rats, but it may have adverse effects on the glucose metabolism. The effect of.2. without calorie intake on intestinal microflora is greater than that of the limit calorie intake..3. drinking water break can turn the bacilli mainly intestinal flora into Poulet Was bacteria, and to pervo. The structure of intestinal microflora with the main bacteria was not much affected by.4.. The body could consume the energy related metabolic pathway during drinking water feeding, consumption of fatty acid, muscle, and nucleic acid to maintain the energy consumption of the body.5.. After drinking water for third days after drinking water, the metabolic products of the body recovered to the normal level, but the intestinal flora was still not. It shows that the metabolic regulation of the body is more sensitive than the intestinal flora on the change of diet. In general, the effect of feeding on the intestinal microflora is more distinct, and the prevention and treatment of clinical diseases still need to be further studied, but it can provide a new perspective for the obese patients.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.2

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