基于宏基因组学的慢性阻塞性肺疾病急性加重患者肠道菌群结构与功能相关性研究

Association between gut microbiota and clinical features in patients with acute exacerbation chronic obstructive pulmonary disease based on metagenomics study

关键词:  慢性阻塞性肺疾病  肠道菌群  宏基因测序

Key words:  chronic obstructive pulmonary disease  gut microbiota  metage-nomics sequencing

摘要:

目的研究慢性阻塞性肺疾病急性加重（acute exacerbation of chronic obstructive pulmonary disease, AECOPD）患者的肠道菌群结构与功能基因、耐药基因、毒力因子的表达差异，探索肠道菌群在AECOPD发生发展中的作用。方法2020年12月—2021年10月，入选符合AECOPD诊断的患者共15例，并选取年龄、吸烟史相匹配的健康对照组5例，收集所有肠道粪便标本，进行宏基因测序，结合临床数据，分析AECOPD与对照组肠道菌群结构及功能等方面的差异性。结果AECOPD患者肠道微生物α多样性与对照组无统计学差异，β多样性存在差异（P=0.022）。AECOPD组栖粪杆菌属（Faecalibacterium）、布劳特氏菌属（Blautia）、厌氧棒菌属（Anaerostipes）相对丰度低于对照组，肠球菌属（Enterococcus）、Lachnoclostridium相对丰度高于对照组（P<0.05），AECOPD信号通路富集在糖酵解Ⅰ、Ⅱ通路，肠球菌属相关的耐药基因及毒力因子表达丰度增加。AECOPD在属水平表达相对丰度较高的肠道微生物菌群为肠球菌属（Enterococcus），其表达相对丰度与年龄呈正相关（P<0.05）；肠球菌属表达丰度与耐药基因msrC、efmA表达丰度呈正相关（P<0.05），与毒力因子acm、EcbA/fss3表达丰度呈正相关（P<0.05）。栖粪杆菌属、布劳特氏菌属、厌氧棒菌属的表达丰度与AECOPD用力肺活量（force vital capacity, FVC）呈正相关（P<0.05）；布劳特氏菌属、厌氧棒菌属的表达丰度与BMI呈正相关（P<0.05）；布劳特氏菌属、厌氧棒菌属两者的表达丰度呈正相关（P<0.05）；Lachnoclostridium属的表达丰度与血液中CD4+/CD8+T淋巴细胞比例呈正相关（P<0.05）。结论AECOPD患者肠球菌属（Enterococcus）相对丰度高，栖粪杆菌属、布劳特氏菌属、厌氧棒菌属相对丰度较低，信号通路富集在糖酵解相关代谢通路。栖粪杆菌属、布劳特氏菌属、厌氧棒菌属的表达丰度与临床指标FVC及T淋巴细胞比例存在一定的相关性，提示肠道微生态的失衡可能通过“肠-肺轴”参与AECOPD的发生发展。

Abstract:

ObjectiveTo investigate the association between gut microbiota and clinical features in patients with acute exacerbation chronic obstructive pulmonary disease(AECOPD) using metagenomics approach. MethodsFifteen patients with AECOPD and 5 healthy controls were enrolled in this study and their feces were collected. The metagenomic sequencing of gut microbiota was performed and compared between AECOPD group and control group. ResultsThere was no significant difference in the alpha diversity of gut microbiota between AECOPD group and healthy controls, while there was significant difference in beta diversity between two groups(P=0.022). Faecalibacterium, Blautia and Anaerostipes were less abundant in AECOPD group, while Enterococcus and Lachnochlostridium were more abundant in AECOPD group compared with healthy controls(P<0.05). Glycolysis Ⅰ and Ⅱ signal pathway were upregulated in AECOPD group. The expression abundance of Enterococcus related antibiotic resistance genes(including msrC and efmA) and virulence factors(including acm and EcbA/fss3) were increased. In patients with AECOPD, the abundance of Enterococcus and Lachnoclostridium showed a positive correlation with age(P<0.05), the abundance of Enterococcus showed a positive correlation with antibiotic resistance genes(msrC and efmA) and virulence factors(acm and EcbA/fss3)(P<0.05). The abundance of Faecalibacterium, Blautia and Anaerostipes showed a positive correlation with force vital capacity(P<0.05), the abundance of Blautia and Anaerostipes showed a positive correlation with body mass index(P<0.05). The abundance of Blautia showed a positive correlation with the abundance of Anaerostipes. The abundance of Faecalibacterium showed a positive correlation with peripheral blood CD4+/CD8+ T lymphocyte ratio(P<0.05). ConclusionThe abundance of Faecalibacterium, Blautia，Anaerostipes，Enterococcus and Lachnoclostridium shows a certain correlation with clinical features, which indicates that intestinal microbial dysbiosis may be involved in the pathogenesis of AECOPD by the gut-lung axis. This study might provide experimental evidence for exploring the mechanism of the gut microbiota on the occurrence and development of AECOPD, and potential targets for prevention and intervention of AECOPD.