海軍長遠(yuǎn)航官兵腸道菌群多樣性研究
發(fā)布時(shí)間：2018-10-05 08:14
【摘要】：研究目的:健康人胃腸(GI)道內(nèi)寄居著種類繁多的微生物,這些微生物統(tǒng)稱為腸道菌群。腸道菌群在腸道微環(huán)境下發(fā)揮著重要作用,參與物質(zhì)代謝,促進(jìn)免疫系統(tǒng)的發(fā)育等。人體各種內(nèi)外環(huán)境的變化會對腸道菌群的種類、結(jié)構(gòu)以及功能產(chǎn)生影響,人體腸道菌群與宿主之間存在著共生關(guān)系,腸道菌群的結(jié)構(gòu)與多樣性改變也會影響其宿主的發(fā)育、免疫、消化與代謝[1,2,3]。通過不斷的適應(yīng)與調(diào)節(jié),菌群、宿主與環(huán)境共同維護(hù)著宿主的生理平衡,有助于維持宿主健康。然而,當(dāng)正常菌群中各菌種間的比例發(fā)生較大幅度變化而超出正常范圍的狀態(tài)時(shí),稱為菌群失調(diào)。目前腸道菌群在腸道相關(guān)疾病的發(fā)生、發(fā)展中的作用及其機(jī)制研究尚不成熟,仍有許多問題亟待解決。除此之外,腸道菌群與肥胖、糖尿病、心梗、抑郁癥等多個(gè)領(lǐng)域的疾病均存在關(guān)聯(lián)[2,3,4]。為了分析長遠(yuǎn)航任務(wù)官兵腸道菌群結(jié)構(gòu)和多樣性特征,本研究著眼于長遠(yuǎn)航任務(wù)中腸道菌群結(jié)構(gòu)的變化對長遠(yuǎn)航官兵健康的意義,尋找應(yīng)對不利變化的措施,預(yù)防疾病的發(fā)生,為提供長遠(yuǎn)航官兵的生活質(zhì)量提供依據(jù)。研究方法:本研究通過使用高通量測序技術(shù),對29名長遠(yuǎn)航任務(wù)官兵于長遠(yuǎn)航出發(fā)前,45天,90天和135天(錨地靠港前)4個(gè)不同時(shí)間節(jié)點(diǎn),將其分為A、B、C、D四組,進(jìn)行糞便樣品采集,提取糞便樣本總DNA;根據(jù)細(xì)菌16S r DNA V3-V5區(qū)設(shè)計(jì)引物進(jìn)行擴(kuò)增,利用Illumina Miseq平臺對腸道菌群DNA進(jìn)行高通量測序;測序結(jié)果經(jīng)過Reads拼接,OTUs(operationaltaxonomic units)聚類,物種注釋,α多樣性分析,最終得到糞便菌群的物種豐度信息。結(jié)果:在116例樣本中,絕大部分reads分屬11個(gè)門,占腸道菌群99%的有5個(gè)門,分別為擬桿菌門(Bacteroidetes)、厚壁菌門(Firmicutes)、變形菌門(Proteobacteria)、梭桿菌門(Fusobacteria)、放線菌門(Actinobacteria)。共測得48屬,排名前10位的屬有:擬桿菌屬(Bacteroides)、普氏菌屬(Prevotella)、毛螺菌屬(Lachnospira)、瘤胃球菌屬(Ruminococcaceae)、Alistipes屬、考拉桿菌屬(Phascolarctobacterium)、副桿菌屬(Parabacteroides)、薩特氏桿菌屬(Sutterella)、羅氏菌屬(Roseburia)、戴阿利斯特菌屬(Dialister)。擬桿菌門(Bacteroidetes)在四組中豐度均為最高,擬桿菌門分別占到60.01%,62.56%,65.05%,57.51%,在116份樣品中所占比例為61.30%;厚壁菌門(Firmicutes)在四組中豐度次之,分別占到34.62%,32.19%,27.21%,37.6%,在116份樣品中所占比例為32.90%;豐度較高的門包括變形菌門、梭桿菌門、放線菌門(Actinobacteria)。此外,四組菌群結(jié)構(gòu)差異性表現(xiàn)為:擬桿菌門(Bacteroidetes)在B組中的豐度高于A組(60.01%vs 62.56%,P0.05),C組的豐度高于B組(65.05%vs 62.56%,P0.05),D組中的豐度低于C組(57.51%vs 65.05%,P0.05),而A組與D組間有明顯差異(60.01%%vs57.51%,P0.05);厚壁菌門(Firmicutes)在A組中的豐度高于B組(34.62%vs 32.19%,P0.05),B組中的豐度高于C組(32.19%vs 27.21%,P0.05),C組中的豐度低于D組(27.21%vs 37.60%,P0.05),而A組與D組間有顯著性差異(34.62%vs 37.60%,P0.05)。α多樣性分析結(jié)構(gòu)顯示,A、D兩組的腸道菌群多樣性高于B、C兩組。說明長遠(yuǎn)航過程中任務(wù)官兵的腸道菌群多樣性不斷降低,于長遠(yuǎn)航結(jié)束時(shí)又大致恢復(fù)到長遠(yuǎn)航前水平。OTUs比對結(jié)果顯示,長遠(yuǎn)航開始后,菌群結(jié)構(gòu)發(fā)生了一些變化:從門的結(jié)構(gòu)上新增了綠彎菌門(Chloroflexi);綱的水平上新增了噬纖維菌綱(Cytophagia),鞘氨醇桿菌綱(Sphingobacteriia),厭氧繩菌綱(Anaerolineae),芽孢桿菌綱(Bacilli);屬的水平上新增了海洋桿菌屬(Pontibacter),脫硫弧菌屬(Desulfovibrio),假單孢菌屬(Preudomonas),熱單孢菌屬(Thermomonas)。同時(shí)也有一些菌屬在長遠(yuǎn)航后消失了,綱的水平上包括Melainabacteria,疣微菌綱(Verrucomicrobiae);屬的水平上包括Hungatella,Marvinbryantia,檸檬酸桿菌屬(Cltrobacter),Cloaclbacillus,疣微菌科的Akkermansia屬。結(jié)論:這些結(jié)果表明,長遠(yuǎn)航過程中任務(wù)官兵在腸道菌群多樣性上與菌群結(jié)構(gòu)均發(fā)生了比較大的變化,但在返航時(shí)其多樣性及菌群結(jié)構(gòu)又逐漸恢復(fù);同時(shí)長遠(yuǎn)航后腸道菌群致病菌數(shù)量多于健康人群,而擬桿菌類有益菌低于健康人群且在長遠(yuǎn)航后任務(wù)官兵的腸道菌群中出現(xiàn)了水生環(huán)境中的菌屬分布。
[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.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R82
本文編號：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.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R82
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 胡紅衛(wèi);黃永坤;;腸道菌群與相關(guān)疾病的研究進(jìn)展[J];中國微生態(tài)學(xué)雜志;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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