海军长远航官兵肠道菌群多样性研究
发布时间：2018-10-05 08:14
【摘要】：研究目的:健康人胃肠(GI)道内寄居着种类繁多的微生物,这些微生物统称为肠道菌群。肠道菌群在肠道微环境下发挥着重要作用,参与物质代谢,促进免疫系统的发育等。人体各种内外环境的变化会对肠道菌群的种类、结构以及功能产生影响,人体肠道菌群与宿主之间存在着共生关系,肠道菌群的结构与多样性改变也会影响其宿主的发育、免疫、消化与代谢[1,2,3]。通过不断的适应与调节,菌群、宿主与环境共同维护着宿主的生理平衡,有助于维持宿主健康。然而,当正常菌群中各菌种间的比例发生较大幅度变化而超出正常范围的状态时,称为菌群失调。目前肠道菌群在肠道相关疾病的发生、发展中的作用及其机制研究尚不成熟,仍有许多问题亟待解决。除此之外,肠道菌群与肥胖、糖尿病、心梗、抑郁症等多个领域的疾病均存在关联[2,3,4]。为了分析长远航任务官兵肠道菌群结构和多样性特征,本研究着眼于长远航任务中肠道菌群结构的变化对长远航官兵健康的意义,寻找应对不利变化的措施,预防疾病的发生,为提供长远航官兵的生活质量提供依据。研究方法:本研究通过使用高通量测序技术,对29名长远航任务官兵于长远航出发前,45天,90天和135天(锚地靠港前)4个不同时间节点,将其分为A、B、C、D四组,进行粪便样品采集,提取粪便样本总DNA;根据细菌16S r DNA V3-V5区设计引物进行扩增,利用Illumina Miseq平台对肠道菌群DNA进行高通量测序;测序结果经过Reads拼接,OTUs(operationaltaxonomic units)聚类,物种注释,α多样性分析,最终得到粪便菌群的物种丰度信息。结果:在116例样本中,绝大部分reads分属11个门,占肠道菌群99%的有5个门,分别为拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、梭杆菌门(Fusobacteria)、放线菌门(Actinobacteria)。共测得48属,排名前10位的属有:拟杆菌属(Bacteroides)、普氏菌属(Prevotella)、毛螺菌属(Lachnospira)、瘤胃球菌属(Ruminococcaceae)、Alistipes属、考拉杆菌属(Phascolarctobacterium)、副杆菌属(Parabacteroides)、萨特氏杆菌属(Sutterella)、罗氏菌属(Roseburia)、戴阿利斯特菌属(Dialister)。拟杆菌门(Bacteroidetes)在四组中丰度均为最高,拟杆菌门分别占到60.01%,62.56%,65.05%,57.51%,在116份样品中所占比例为61.30%;厚壁菌门(Firmicutes)在四组中丰度次之,分别占到34.62%,32.19%,27.21%,37.6%,在116份样品中所占比例为32.90%;丰度较高的门包括变形菌门、梭杆菌门、放线菌门(Actinobacteria)。此外,四组菌群结构差异性表现为:拟杆菌门(Bacteroidetes)在B组中的丰度高于A组(60.01%vs 62.56%,P0.05),C组的丰度高于B组(65.05%vs 62.56%,P0.05),D组中的丰度低于C组(57.51%vs 65.05%,P0.05),而A组与D组间有明显差异(60.01%%vs57.51%,P0.05);厚壁菌门(Firmicutes)在A组中的丰度高于B组(34.62%vs 32.19%,P0.05),B组中的丰度高于C组(32.19%vs 27.21%,P0.05),C组中的丰度低于D组(27.21%vs 37.60%,P0.05),而A组与D组间有显著性差异(34.62%vs 37.60%,P0.05)。α多样性分析结构显示,A、D两组的肠道菌群多样性高于B、C两组。说明长远航过程中任务官兵的肠道菌群多样性不断降低,于长远航结束时又大致恢复到长远航前水平。OTUs比对结果显示,长远航开始后,菌群结构发生了一些变化:从门的结构上新增了绿弯菌门(Chloroflexi);纲的水平上新增了噬纤维菌纲(Cytophagia),鞘氨醇杆菌纲(Sphingobacteriia),厌氧绳菌纲(Anaerolineae),芽孢杆菌纲(Bacilli);属的水平上新增了海洋杆菌属(Pontibacter),脱硫弧菌属(Desulfovibrio),假单孢菌属(Preudomonas),热单孢菌属(Thermomonas)。同时也有一些菌属在长远航后消失了,纲的水平上包括Melainabacteria,疣微菌纲(Verrucomicrobiae);属的水平上包括Hungatella,Marvinbryantia,柠檬酸杆菌属(Cltrobacter),Cloaclbacillus,疣微菌科的Akkermansia属。结论:这些结果表明,长远航过程中任务官兵在肠道菌群多样性上与菌群结构均发生了比较大的变化,但在返航时其多样性及菌群结构又逐渐恢复;同时长远航后肠道菌群致病菌数量多于健康人群,而拟杆菌类有益菌低于健康人群且在长远航后任务官兵的肠道菌群中出现了水生环境中的菌属分布。
[Abstract]:Objective: The gastrointestinal (GI) tract of a healthy person is a large number of microorganisms, which are collectively referred to as intestinal flora. The intestinal flora plays an important role in intestinal microenvironment, participates in substance metabolism, promotes the development of the immune system, and so on. The variety, structure and function of intestinal flora were affected by various changes of internal and external environment of human body. There was a symbiotic relationship between intestinal flora and host, and the structure and diversity of intestinal flora affected its host development, immunity, digestion and metabolism[1, 2, 3]. The physiological balance of the host is maintained by continuous adaptation and regulation, flora, host and environment, which helps to maintain the host's health. However, when the proportion of each strain in the normal bacteria population varies considerably beyond the normal range, it is called a flora imbalance. At present, there are still many problems to be solved in intestinal flora in intestinal tract related diseases, its role in development and its mechanism. In addition, intestinal flora is associated with diseases such as obesity, diabetes, depression, depression, etc.[2, 3, 4]. in ord to analyze that structure and diversity of intestinal flora in long-distance voyage mission, this study aims at the significance of the change of intestinal flora structure in long-distance voyage mission to the health of long-distance voyage officers and men, to find measures to cope with adverse changes, and to prevent the occurrence of diseases. In order to provide the basis for providing the quality of life of long-distance officers and soldiers. Methods: Through the use of high-flux sequencing technology, 29 long-distance pilots were divided into four groups: A, B, C and D before long voyage, 45 days, 90 days and 135 days (before arrival). The total DNA of the stool sample is extracted; the primers are designed according to the design primers of the bacterial 16S rDNA V3-V5 region to carry out high-flux sequencing on the intestinal flora DNA by using the Illumina Miseq platform; the sequencing result is spliced by a Reads; the OTOMY (Solutionaltaxonomics units) poly, the species annotation, the species diversity analysis, and finally obtaining the species abundance information of the fecal bacteria population. Results: Among 116 samples, the majority of reads belong to 11 doors, accounting for 99% of the intestinal flora, 5 of which are Bacteroidites, Firmicutes, Proteobacteria, Clostridium and Actinobacteria. A total of 48 genera, 10 of the top 10 genera are Bacteroides, Prevotella, Lachnospira, Ruminococaceae, Alfulpes, Phascolcobacterium, Parbacteroides, Suttertella, Roseuria, Dialister. The abundance of Bacteroidites in four groups was the highest, and the proposed bacteria accounted for 60. 01%, 62. 56%, 65. 05%, 57. 51% respectively. Among 116 samples, the proportion was 66.1%, and the abundance of thick wall bacteria in four groups was 34.62%, 32.19%, 27.21%, 37.6%, respectively. Among 116 samples, the proportion was 32.90%, and the gates with higher abundance include deformation bacteria gate, shuttle gate, actinomyces door (Acinetobacter). In the group B, the abundance of Bacteroidites in group B was higher than that in group A (60. 01% vs 62 56%, P0.05), and the abundance of group C was higher than that in group B (65. 05% vs 62 56%, P0.05), and the abundance in group D was lower than that in group C (57. 51% vs 65. 05%, P0.05). The abundance in group A was higher than that in group B (34.62% vs 32.19%, P0.05). The abundance in group B was higher than that in group C (32.19% vs 27.21%, P0.05), and the abundance in group C was lower than group D (27.21% vs 37.60%, P0.05). There was significant difference between group A and group D (34.62% vs 37.60%, P0.05). The diversity analysis showed that the diversity of intestinal flora in group A and D was higher than that of group B and C. The diversity of the intestinal flora of the officers and soldiers in the long-distance voyage was reduced, and the long-distance voyage was resumed at the end of the long voyage. The results showed that after long voyage, there were some changes in the structure of bacteria group: from the structure of the door, the bacterium group structure was added with the Chloroplast; the level of the class increased the Cytophidia, Sphingobacter, Anaerelinae and Bacilli; The genus Pontibacter, Desulfovibrio, Preudus and Thermomonas are newly added at the level of genera. At the same time, some fungi disappeared after long voyage, including Melainabacterodia, Verrucomicrobacter; belonging to the genus Herungatella, Marybryantia, Clticrobacter, Cloaca, and Akamamansia of the Microbacterium family. Conclusion: These results show that the diversity and flora structure of the mission officers and soldiers in the long voyage are relatively large, but the number of pathogenic bacteria in the intestinal flora after long voyage is more than that of healthy people. The bacteria in the aquatic environment were found to be lower than healthy people and in the intestinal flora of the officers and soldiers after long voyage.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R82
本文编号：2252654
[Abstract]:Objective: The gastrointestinal (GI) tract of a healthy person is a large number of microorganisms, which are collectively referred to as intestinal flora. The intestinal flora plays an important role in intestinal microenvironment, participates in substance metabolism, promotes the development of the immune system, and so on. The variety, structure and function of intestinal flora were affected by various changes of internal and external environment of human body. There was a symbiotic relationship between intestinal flora and host, and the structure and diversity of intestinal flora affected its host development, immunity, digestion and metabolism[1, 2, 3]. The physiological balance of the host is maintained by continuous adaptation and regulation, flora, host and environment, which helps to maintain the host's health. However, when the proportion of each strain in the normal bacteria population varies considerably beyond the normal range, it is called a flora imbalance. At present, there are still many problems to be solved in intestinal flora in intestinal tract related diseases, its role in development and its mechanism. In addition, intestinal flora is associated with diseases such as obesity, diabetes, depression, depression, etc.[2, 3, 4]. in ord to analyze that structure and diversity of intestinal flora in long-distance voyage mission, this study aims at the significance of the change of intestinal flora structure in long-distance voyage mission to the health of long-distance voyage officers and men, to find measures to cope with adverse changes, and to prevent the occurrence of diseases. In order to provide the basis for providing the quality of life of long-distance officers and soldiers. Methods: Through the use of high-flux sequencing technology, 29 long-distance pilots were divided into four groups: A, B, C and D before long voyage, 45 days, 90 days and 135 days (before arrival). The total DNA of the stool sample is extracted; the primers are designed according to the design primers of the bacterial 16S rDNA V3-V5 region to carry out high-flux sequencing on the intestinal flora DNA by using the Illumina Miseq platform; the sequencing result is spliced by a Reads; the OTOMY (Solutionaltaxonomics units) poly, the species annotation, the species diversity analysis, and finally obtaining the species abundance information of the fecal bacteria population. Results: Among 116 samples, the majority of reads belong to 11 doors, accounting for 99% of the intestinal flora, 5 of which are Bacteroidites, Firmicutes, Proteobacteria, Clostridium and Actinobacteria. A total of 48 genera, 10 of the top 10 genera are Bacteroides, Prevotella, Lachnospira, Ruminococaceae, Alfulpes, Phascolcobacterium, Parbacteroides, Suttertella, Roseuria, Dialister. The abundance of Bacteroidites in four groups was the highest, and the proposed bacteria accounted for 60. 01%, 62. 56%, 65. 05%, 57. 51% respectively. Among 116 samples, the proportion was 66.1%, and the abundance of thick wall bacteria in four groups was 34.62%, 32.19%, 27.21%, 37.6%, respectively. Among 116 samples, the proportion was 32.90%, and the gates with higher abundance include deformation bacteria gate, shuttle gate, actinomyces door (Acinetobacter). In the group B, the abundance of Bacteroidites in group B was higher than that in group A (60. 01% vs 62 56%, P0.05), and the abundance of group C was higher than that in group B (65. 05% vs 62 56%, P0.05), and the abundance in group D was lower than that in group C (57. 51% vs 65. 05%, P0.05). The abundance in group A was higher than that in group B (34.62% vs 32.19%, P0.05). The abundance in group B was higher than that in group C (32.19% vs 27.21%, P0.05), and the abundance in group C was lower than group D (27.21% vs 37.60%, P0.05). There was significant difference between group A and group D (34.62% vs 37.60%, P0.05). The diversity analysis showed that the diversity of intestinal flora in group A and D was higher than that of group B and C. The diversity of the intestinal flora of the officers and soldiers in the long-distance voyage was reduced, and the long-distance voyage was resumed at the end of the long voyage. The results showed that after long voyage, there were some changes in the structure of bacteria group: from the structure of the door, the bacterium group structure was added with the Chloroplast; the level of the class increased the Cytophidia, Sphingobacter, Anaerelinae and Bacilli; The genus Pontibacter, Desulfovibrio, Preudus and Thermomonas are newly added at the level of genera. At the same time, some fungi disappeared after long voyage, including Melainabacterodia, Verrucomicrobacter; belonging to the genus Herungatella, Marybryantia, Clticrobacter, Cloaca, and Akamamansia of the Microbacterium family. Conclusion: These results show that the diversity and flora structure of the mission officers and soldiers in the long voyage are relatively large, but the number of pathogenic bacteria in the intestinal flora after long voyage is more than that of healthy people. The bacteria in the aquatic environment were found to be lower than healthy people and in the intestinal flora of the officers and soldiers after long voyage.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R82
【参考文献】
相关期刊论文 前2条
1 胡红卫;黄永坤;;肠道菌群与相关疾病的研究进展[J];中国微生态学杂志;2017年01期
2 Jennifer K Beatty;Amol Bhargava;Andre G Buret;;Post-infectious irritable bowel syndrome:Mechanistic insights into chronic disturbances following enteric infection[J];World Journal of Gastroenterology;2014年14期
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