Association study of gut flora in Wilson's disease through high-throughput sequencing.
Association study of gut flora in Wilson's disease through high-throughput sequencing.
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通过高通量测序进行威尔逊病肠道菌群的关联研究
DOI：
10.1097/md.0000000000011743
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发表时间：
2018-08
期刊：
影响因子：
1.6
通讯作者：
Cheng N
中科院分区：
文献类型：
--
作者：
Geng H;Shu S;Dong J;Li H;Xu C;Han Y;Hu J;Han Y;Yang R;Cheng N
摘要 本研究通过高通量测序技术分析威尔逊病（WD）患者与健康人群肠道菌群多态性的差异，探讨WD与肠道菌群多态性的相关性。共招募 22 例 WD 患者和 22 名健康人作为对照。提取所有受试者粪便标本中的总DNA，扩增16S rRNA基因V4高可变区，并通过高通量测序进行测序。测序结果通过&agr;进行分析。多样性和 &bgr;多样性。计算未加权UniFrac距离矩阵并通过算术平均的未加权配对组法(UPGMA)构建树。数据优化后共获得2,548,262条序列，WD组平均序列数为36,836±4104，正常对照组为35,051±3075，两组平均序列数无显着差异。 OTU分析显示，WD组获得2663个OTU，对照组获得3271个OTU，其中941个为普通OTU。菌落多样性分析显示，WD组和对照组肠道菌群分属5个门，分别为拟杆菌门、厚壁菌门、变形菌门、梭杆菌门和软壁菌门。 WD组中拟杆菌门丰度显着低于对照组（67.19% vs 76.75%，P < .001），厚壁菌门丰度显着低于对照组（26.18% vs 19.83%，P < .001），变形菌门丰度显着低于对照组（4.31% vs 3.09%， P < .05)、梭杆菌(1.88% vs 0.04%, P < .001)显着高于对照组。与对照组相比，属水平上拟杆菌丰度（4.85% vs 4.6%，P < .05）、粪杆菌丰度（2.92% vs 2.13%，P < .05）、巨藻丰度（0.84% vs 0.22%，P < .001）、毛螺菌丰度WD组（0.16% vs 0.09%，P < .001）显着增加，而普雷沃氏菌（1.63% vs 2.48%，P < .001）、罗斯布里亚菌（0.75% vs 1.39%，P < .001）和棕囊菌属（1.72% vs 1.72%）丰度显着增加。 2.45%, P < .001) WD组显着下降。 PCoA和UPGMA树分析显示，两组之间的肠道微生物组成存在显着差异。 WD患者肠道菌群多样性和组成显着低于健康对照，肠道菌群多样性可能与WD的存在有关。
Abstract In this study, we analyzed the difference of intestinal flora polymorphisms between Wilson's disease (WD) patients and healthy people by high-throughput sequencing technology, and explored the correlation between WD and intestinal flora polymorphism. A total of 22 cases of WD patients and 22 healthy persons as control were recruited. The total DNA was extracted from the fecal specimens of all the subjects, V4 high variable region of 16S rRNA gene was amplified and sequenced by high-throughput sequencing. The sequencing results were analyzed by &agr; diversity and &bgr; diversity. The unweighted UniFrac distance matrices were calculated and trees were built by unweighted-pair group method with arithmetic mean (UPGMA). A total of 2,548,262 sequences were obtained after the data are optimized, the average sequences in the WD group was 36,836 ± 4104 and it was 35,051 ± 3075 in the normal control group, there was no significant difference in the average sequence number between the 2 groups. OTU analysis showed that 2663 OTU were obtained in WD group, and 3271 OTU were obtained in the control group, of which 941 were common OTU. Colony diversity analysis showed that the intestinal flora of WD group and control group belonged to 5 phyla, they were Bacteroidetes, Firmicutes, Proteobacteria, Fusobacteria, and Tenericutes, respectively. In WD group, the abundance of Bacteroidetes was significantly lower than that of the control group (67.19% vs 76.75%, P < .001), and the abundance of Firmicutes (26.18% vs 19.83%, P < .001), Proteobacteria (4.31% vs 3.09%, P < .05), Fusobacteria (1.88% vs 0.04%, P < .001) were significantly higher than that of control group. Compared with the control group at the level of the genus, the abundance of Bacteroides (4.85% vs 4.6%, P < .05), Faecalibacterium (2.92% vs 2.13%, P < .05), Megamonas (0.84% vs 0.22%, P < .001), Lachnospira (0.16% vs 0.09%, P < .001) significantly increased in WD group, while the abundance of Prevotella (1.63% vs 2.48%, P < .001), Roseburia (0.75% vs 1.39%, P < .001) and Phascolarctobacterium (1.72% vs 2.45%, P < .001) significantly decreased in WD group. PCoA and UPGMA tree analysis showed that there were significant differences of gut microbial compositions between the 2 groups. The diversity and composition of intestinal flora in the WD patients were significantly lower than those in the healthy controls, and the diversity of intestinal flora may be associated with the presence of WD.