宏基因组二代测序比较慢性阻塞性肺疾病稳定期患者与健康人的肠道菌群差异

Comparison of gut microflora between stable chronic obstructive pulmonary disease patients and healthy controls by metagenomic second generation sequencing

目的分析健康人与慢性阻塞性肺疾病(COPD)稳定期用药和未用药患者的肠道菌群相对丰度及功能基因差异。方法本研究为病例对照研究。采用非随机抽样方法,收集2019年1月至2019年12月在广州医科大学附属第一医院就诊的体检健康人10名(A组)、COPD稳定期未用药患者10例(B组)和COPD稳定期用药患者10例(C组)的粪便,收集3组患者一般资料,采用宏基因组二代测序方法比较3组肠道相对丰度及功能基因。结果研究对象的年龄、身高、体质量、体质量指数差异无统计学意义(P值均>0.05)。基于相似性分析与主成分分析,3组间差异无统计学意义;基于Metastats差异分析,与A组比较,B组的普氏菌属、马赛普菌属和巨球型菌属均显著减少(P值均<0.05);而C组的埃希菌属显著增加(P=0.045),C组的马赛普菌属和巨球型菌属显著减少(P值均<0.05);B组与C组间差异无统计学意义(P>0.05)。基于功能基因分析,在level3水平上,与A组相比,B组的果糖和甘露糖代谢、萜类化合物生物合成、氨基酸合成、同源重组、淀粉和蔗糖代谢功能基因通路显著减少,细菌双组分调节系统通路显著增加;而C组的果糖和甘露糖代谢、萜类化合物生物合成、氨基酸合成、同源重组功能基因通路显著减少,细菌分泌系统、细菌双组分调节系统、糖酵解与合成显著增加(P值均<0.05);B组与C组比较差异无统计学意义(P>0.05)。结论与健康人比较,COPD稳定期患者的肠道菌群存在特征性改变,复诊治疗患者与初诊未用药治疗患者间的差异无统计学意义;COPD稳定期患者的果糖和甘露糖代谢、萜类化合物生物合成、氨基酸合成和同源重组功能基因通路显著减少,提示能量代谢功能下降。

Objective To analyze the relative abundance and functional genes of intestinal microflora in healthy subjects and patients with chronic obstructive pulmonary disease(COPD).Methods This was a case-control study.Non-random sampling was used.The feces of 10 healthy people(Group A),10 patients with COPD who received no medication(Group B),and 10 patients with COPD who received medication(Group C)in the First Affiliated Hospital of Guangzhou Medical University from January 2019 to December 2019 were collected.Metagenomic second generation sequencing was used to compare the relative abundance and functional genes in the three groups.Results There were no significant differences in age,height,weight,and BMI(all P>0.05).Based on similarity analysis and principal component analysis,there was no significant difference among the three groups.Based on the difference analysis of Metastats,Prevotella,Prevotellamassilia,and Megasera in Group B were significantly lower than those in Group A(all P<0.05).However,in Group C the number of Escherichia(P=0.045)was significantly increased and the numbers of Prevotellamassia and Megasphaera were significantly decreased(all P<0.05),with no significant difference between Group B and C(P>0.05).Based on functional gene analysis,at level 3,compared with Group A,the functional gene pathways of ko00051 fructose and mannose metabolism,ko00900 terpenoid backbone biosynthesis,ko01230 biosynthesis of amino acids,ko03440 homologous recombination,and ko00500 starch and sucrose metabolism were significantly reduced,ko02020 two-component system pathway was significantly increased in Group B,and the functional gene pathways of ko00051 fructose and mannose metabolism,ko00900 terpenoid backbone biosynthesis,ko01230 biosynthesis of amino acids,and ko03440 homologous recombination reduced significantly,ko03070 bacterial secretion system,ko02020 two-component system,and ko00010 glycolysis/gluconeogenesis increased significantly in Group C(all P<0.05),with no statistical significance between Group B and C(P>0.05).Conclusions Compared with healthy subjects,the gut microflora of COPD patients in stable stage had characteristic changes,and there was no significant difference between the patients who received follow-up treatment and the patients who didn′t.Fructose and mannose metabolism,terpenoid biosynthesis,amino acid synthesis,and homologous recombination functional gene pathway were significantly reduced in patients with stable COPD,indicating that energy metabolism function was decreased.