广西巴马长寿老人肠道菌群及其与膳食纤维多糖饮食关联性研究

发布时间：2018-03-06 10:05

  本文选题：广西巴马　切入点：长寿老人　出处：《广西大学》2015年博士论文　论文类型：学位论文

【摘要】：膳食纤维是多糖的重要组成成分,虽其本身不能被宿主消化吸收利用,但对胃肠功能、其他重要营养素和宿主肠道菌群的平衡都具有重要作用,也是保证人体健康的重要物质之一。广西巴马是世界著名的长寿之乡,从其饮食结构研究入手,探索膳食纤维多糖饮食和广西巴马长寿老人肠道菌群相关性,对深入了解膳食纤维多糖功效,阐明饮食与健康长寿关系,服务社会具有重要的意义。目前,世界各个地区长寿老人肠道菌群特征正在不断被报道。然而,对于广西巴马这样一个世界著名的长寿乡,历史悠久且生活饮食独具特色的人群,迄今为止仍没有关于该人群肠道菌群与其饮食结构相关性的研究报道。本研究利用流行病学饮食调查问卷FFQ23、Illumina MiSeq高通量测序技术、变性梯度凝胶电泳(PCR-DGGE)、属特异性克隆文库和种特异性荧光定量PCR技术,对广西巴马百岁老人、巴马80-99岁老年人和南宁80-99岁老年人饮食情况、肠道菌群结构、双歧杆菌群落结构、乳酸菌群落结构进行探索研究,旨在揭示广西巴马长寿老人肠道菌群结构特征,深入分析饮食结构与巴马长寿老人肠道菌群的密切联系。饮食调查结果表明,广西巴马长寿地区饮食特点是以粥类、粗杂粮类、深色蔬菜类、家畜肉类食物摄入为主的膳食纤维多糖饮食模式。与《中国居民膳食指南》推荐量相比,除百岁老人饮食中谷物摄入量达标率为71.43%和巴马80-99岁老年人蔬菜摄入量达标率为52.63%外,长寿地区老年人饮食模式中其他食物摄入量达标率均小于15%,其中畜禽肉类摄入水平显著高于非长寿地区老年人(p0.05)。营养结构分析显示长寿地区膳食纤维摄入量显著高于非长寿地区老人(p0.05),且相比于2004年中国居民膳食营养素参考摄入量,长寿地区百岁老人饮食结构中其他营养素达标率很低的情况下,仅维生素A摄入量达标率为100%。百岁老人三大营养素供能比为：蛋白质17.94%,脂肪35.82%,碳水化合物52.23%,结果显示我国居民膳食指南的推荐量不适合健康高龄老人的饮食需求。基于16S rDNA的Illumina MiSeq高通量测序技术结合多元统计分析识别老年人肠道菌群与其饮食结构、食物摄入量及营养素摄入量之间的相关关系。结果表明,对比80-99岁老年人,巴马百岁老人肠道菌群多样性增加(p0.05)。基于肠道菌群α和β多样性分析结果证实了三组老年人肠道菌群构成差异明显,其中在肠道菌群属水平中罗氏菌属(Roseburia)和埃希氏菌属(Escherichia)的相对丰度在百岁老人粪便中显著偏高(p0.05),而乳酸菌属(Lactobacillus)、栖粪杆菌属(Faecalibacterium)、副拟杆菌属(Parabacteroides)、丁酸弧菌属(Butyricimonas)、粪球菌属(Coprococcus)、巨单胞菌属(Megamonas)、光冈菌属(Mitsuokella)、萨顿氏菌属(Sutterella)和Akkermansia的相对丰度显著偏低(p0.05)。通过与饮食结构、食物摄入量、营养素摄入量相关性分析发现,瘤胃菌科(Ruminococcaceae)与膳食纤维多糖饮食、罗氏菌属(Roseburia)与粗杂粮摄入量和膳食纤维多糖营养素摄入量呈现正相关关系。基于巢式PCR-DGGE.属特异性克隆文库和种特异性荧光定量PCR技术结合多元统计分析识别老年人肠道双歧杆菌群落结构与其饮食结构中的食物摄入量及营养素摄入量之间的相关性。结果表明,百岁老人粪便双歧杆菌群落结构多样性增加。除最小双歧杆菌(Bifidobacterium minimum)、鸡双歧杆菌(B. saecularmay/B. pullorum/B. gallinarum)和摩恩格里艾恩斯双歧杆菌(B. mongoliense)是百岁老年组独有菌种外,齿双歧杆菌(B. dentium)、长双歧杆菌(B. longum)、嗜热双歧杆菌(B. thermophilum)、假链状双歧杆菌／链状双歧杆菌(B. pseudocatenulatum/B. catenulatum)和青春双歧杆菌(B. adolescentis)是三组老年人共有的肠道双歧杆菌。它们的差异性主要表现为：对比巴马80-99岁组,巴马百岁组中长双歧杆菌(B. longum)和齿双歧杆菌(B. dentium)的数量显著增加(p0.05)；对比南宁80-99岁,百岁组中青春双歧杆菌(B. adolescentis)的检出率显著降低(p0.05)。与饮食关联性研究发现,长双歧杆菌(B. longum)的含量与家畜肉类摄入量呈显著负相关(p0.05),青春双歧杆菌(B. adolescentis)的含量与河鲜类摄入量呈显著正相关(p0.05),嗜热双歧杆菌(B. thermophilum)的含量与碳水化合物摄入量呈显著正相关(p0.05)。基于巢式PCR-DGGE、属特异性克隆文库和种特异性荧光定量PCR技术结合多元统计分析手段识别老年人肠道乳酸菌群落结构与其饮食结构中的食物摄入量及营养素摄入量之间的相关关系。结果表明,百岁组粪便乳酸菌群落结构多样性增加,同型发酵乳酸菌数量和检出率增加,而异型发酵乳酸菌群落结构发生重排。它们之间主要的差异表现为：百岁组中嗜酸乳杆菌(L. acidophilus)、发酵乳杆菌(L. fermentum)、融合魏斯氏菌(W. confusa)和乳明串珠菌(L. lactis)的数量显著提高(p0.05),且对比80-99岁组,发酵乳杆菌(L. fermentum)和融合魏斯氏菌(W. confusa)的检测率也显著增强(p0.05)。造成该差异的主要菌种为：融合魏斯氏菌(W. confusa)、嗜酸乳杆菌(L. acidophilus)和罗伊氏乳杆菌(L. reuteri),其中融合魏斯氏菌(W. confusa)与罗伊氏乳杆菌(L. reuteri)存在显著负相关关系。与饮食的关联性研究发现,融合魏斯氏菌(W. confusa)与粥类、粗杂粮及家畜肉类的食物摄入量具有显著正相关关系(p0.05),而罗伊氏乳杆菌(L. reuteri)与膳食纤维摄入量具有显著负相关关系(p0.05)。综上所述,在百岁老人饮食结构中粗杂粮所占的比例高,而他们的肠道菌群中有益菌的数量也高,如罗氏菌属(Roseburia),且益生菌——双歧杆菌和乳酸菌群落多样性都更加丰富。上述的研究结果为健康长寿老人肠道菌群与其膳食纤维多糖饮食关联性学说提供了新的理论依据。
[Abstract]:Dietary fiber is an important component of polysaccharide, although itself cannot be absorbed by the host digestion, but on gastrointestinal function and other important nutrients and gut microflora balance plays an important role, is also one of the most important substances to human health. Guangxi Bama is the world famous Changshouzhixiang, starting from the to explore the structure of food, dietary fiber polysaccharide diet and Guangxi Bama longevity of the elderly intestinal flora correlation, in-depth understanding of the dietary fiber polysaccharide effect, clarify the relationship between health and longevity diet, plays an important role in social services. At present, all regions of the world longevity elderly intestinal flora characteristics are reported. However, for Guangxi Bama such a world famous longevity Township, has a long history and unique life eating people, so far still not on the human intestinal flora and its food structure The Research Report of the questionnaire. The epidemiology of FFQ23 diet used in this study, Illumina MiSeq high-throughput sequencing, denaturing gradient gel electrophoresis (PCR-DGGE), a specific clone library and species-specific fluorescent quantitative PCR technology of Guangxi Bama centenarians, 80-99 year old Nanning Bama and 80-99 year old diet, the structure of intestinal flora, Bifidobacterium community structure, explore and study the community structure of lactic acid bacteria, to reveal the Guangxi Bama longevity of the elderly microflora characteristics, in-depth analysis of diet and Bama longevity of the elderly intestinal flora closely. Dietary survey results show that Guangxi Bama longevity region is characterized by eating porridge, coarse grains dark colored vegetables, livestock, meat food intake of dietary fiber polysaccharide dietary patterns. Compared with the dietary guidelines > < China recommendations, in addition to centenarians diet Valley Intake compliance rate was 71.43% and 80-99 years old in Bama vegetable intake compliance rate was 52.63%, the diet pattern for the elderly longevity areas in the standard rate of other food intake were less than 15%, the meat intake level was significantly higher than that in non elderly longevity area (P0.05). The analysis showed that the longevity area of dietary fiber intake was significantly higher than that of non longevity area the elderly nutrition structure (P0.05), and compared to the dietary reference intake in 2004 China residents, longevity of centenarians in the diet of other nutrients compliance rate is very low, only the standard rate of vitamin A intake of centenarians 100%. energy supply of three major nutrients: protein 17.94%, fat 35.82%. Results showed that 52.23% carbohydrates, dietary requirements recommended dietary guidelines of residents in our country is not suitable for the elderly health. 16S rDNA Illumina MiSeq based on Qualcomm The amount of sequencing technology combined with multivariate statistical analysis to identify elderly intestinal flora and dietary structure, the relationship between food intake and nutrient intake. The results showed that compared to 80-99 years of age or older, Bama centenarians intestinal flora diversity increased (P0.05). The intestinal flora of alpha and beta diversity analysis results confirmed that three group of elderly intestinal flora composition differences based on the intestinal microflora in the genus level of Rothia (Roseburia) and Escherichia coli (Escherichia) is higher significantly in the relative abundance of centenarians in feces (P0.05), and lactic acid bacteria (Lactobacillus), fecal bacteria of the genus habitat (Faecalibacterium), vice Bacteroides (Parabacteroides), butyrivibrio (Butyricimonas), coprococcus (Coprococcus), giant Aeromonas (Megamonas), Mitsuoka sp. (Mitsuokella), Sutton's genus (Sutterella) and the relative abundance of Akkermansia significantly The low (P0.05). With the diet, food intake, correlation analysis of nutrient intake, rumen bacteria, (Ruminococcaceae) and dietary fiber polysaccharide diet, Rothia (Roseburia) showed positive correlation with coarse food intake and nutrient intake of dietary fiber polysaccharide. Nested PCR-DGGE. genus specific clone library and a specific fluorescent quantitative PCR technique combined with multivariate statistical analysis the correlation between food intake and nutritional identification of elderly intestinal Bifidobacterium community structure and diet based on the intakes. The results indicate that the centenarians fecal Bifidobacterium diversity increased. In addition to the minimum of Bifidobacterium (Bifidobacterium minimum), chicken Bifidobacterium (B. saecularmay/B. pullorum/B. gallinarum) and Moen Gerry Irons Bifidobacterium (B. mongoliense) is a hundred years old group unique species, Bifidobacterium breve (B. dentium), Bifidobacterium longum (B. longum), Bifidobacterium thermophilum (B. thermophilum), Bifidobacterium pseudocatenulatum / Bifidobacterium catenulatum (B. pseudocatenulatum/B. catenulatum) and B.adolescentis (B. adolescentis) is a group of three elderly common intestinal bifidobacteria. Their difference is mainly Obama: comparison of 80-99 years old group, Bifidobacterium longum centenarian in Bama (B. longum) and Bifidobacterium breve (B. dentium) significantly increased the number (P0.05); compared with Nanning's 80-99 year old youth, Bifidobacterium centenarian in bacteria (B. adolescentis) the detection rate was significantly lower (P0.05) discovery and research. Association of diet, Bifidobacterium longum (B. longum) and the content of livestock meat intake was negatively correlated (P0.05), B.adolescentis (B. adolescentis) and the content of fish intake is positively related (P0.05), Bifidobacterium thermophilum (B. t Hermophilum) and the content of carbohydrate intake was positively correlated (P0.05). Based on the nested PCR-DGGE, genus specific clone library and species-specific fluorescent quantitative PCR technique combined with multivariate statistical analysis on the relationship between food intake and nutritional means of identification of elderly intestinal lactic acid bacteria community structure and the structure of the food intakes. The results showed that hundred fecal lactic acid bacteria community structure diversity increased, the number of the same type of fermentation of lactic acid bacteria and detection rate increased, while the heterofermentative lactic bacteria community structure rearrangement. The main difference between them is: Lactobacillus acidophilus 100 group (L. acidophilus), l.fermentum (L. fermentum), fusion Wei Adams (W. confusa) and Salmonella Leuconostoc lactis (L. lactis) the number increased significantly (P0.05), and the contrast group of 80-99 years old, l.fermentum (L. fermentum) and weissella strains (W. conf USA) the detection rate also significantly increased (P0.05). The difference is mainly caused by bacteria: weissella strains (W., confusa) (L. acidophilus) of Lactobacillus acidophilus and Lactobacillus reuteri (L. reuteri), which weissella strains (W. confusa) and Lactobacillus reuteri (L. reuteri) there are significant negative correlation. The discovery and study of relevance of the diet, weissella strains (W. confusa) and cereals, grains and livestock meat food intake has a significant positive correlation (P0.05), and Lactobacillus reuteri (L. reuteri) has a significant negative correlation with the intake of dietary fiber (P0.05). To sum up, in the centenarians diet of coarse grains and the proportion of high, and their intestinal flora in bacteria quantity is high, such as Roche sp. (Roseburia), and the probiotic Bifidobacterium and lactic acid bacteria community diversity is more abundant. The above research The results provided a new theoretical basis for the relationship between the intestinal microflora and the dietary fiber polysaccharide diet.

【学位授予单位】：广西大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R153.3

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