尘肺病患者下呼吸道菌群结构特征研究

Structural characteristics of lower respiratory tract microflora in patients with pneumoconiosis

目的探索尘肺病患者及接尘者下呼吸道菌群组成,进一步比较分析两者菌群差异及菌群间的关联。方法于2020年5月至2021年1月,通过前瞻性多中心横断面研究,选取同期就诊于联勤保障部队第九二〇医院呼吸与危重症医学科和昆明市同仁医院呼吸内科,行支气管肺泡灌洗治疗的尘肺病患者及接尘者人群。共纳入24例尘肺病患者(尘肺病组),以16例接尘者(接尘组)为对照,收集2组患者肺泡灌洗液,利用16SrRNA基因V3-V4测序技术及生物信息分析平台,测定并分析其微生物结构组成差异及菌群间的关联。结果与接尘组比较,尘肺病组患者肺泡灌洗液门水平菌群丰度排名前5的菌门相同,依次为:变形菌门、厚壁菌门、拟杆菌门、梭杆菌门、放线菌门。与接尘组比较,尘肺病组患者属水平菌群丰度排名前5的菌属不同,接尘组分别为:假单胞菌属、普氏菌属、链球菌属、无色杆菌属、奈瑟氏菌属,尘肺病组分别为:假单胞菌属、无色杆菌属、链球菌属、罗尔斯通菌属、普氏菌属。Alpha多样性分析结果显示,与接尘组比较,尘肺病组患者菌群多样性有差异(P<0.05),菌群均匀度差异无统计学意义(P>0.05);β多样性显示,两组间菌群结构差异有统计学意义(P<0.05)。单因素菌群关联网络分析示,在尘肺病组及接尘组厚壁菌门和拟杆菌门与其他物种之间相关性较高,呈正相关;变形菌门与其他物种之间相关性较高,呈负相关。结论尘肺病患者及接尘者下呼吸道菌群结构及相对丰度不同,尘肺病患者下呼吸道菌群多样性增加,差异可能与疾病状态有关。

Objective To explore the composition of bacteria in lower respiratory tract of patients with pneumoconiosis and dust exposure,and to compare and analyze the difference and correlation between them.Methods From May 2020 to January 2021,a prospective multicenter cross-sectional study was conducted to select patients with pneumoconiosis who underwent bronchoalveolar lavage treatment at the Respiratory and Critical Care Medical Department of the 920th Hospital of the Joint Support Force and the Respiratory Department of Tongren Hospital in Kunming,as well as the population of dust recipients.A total of 24 patients with pneumoconiosis(pneumoconiosis group)were included,and 16 dust exposed individuals(dust exposed group)were used as controls.Two groups of patients'alveolar lavage fluid were collected.The 16SrRNA gene V3-V4 sequencing technology and bioinformatics analysis platform were used to measure and analyze the differences in microbial structure composition and associations between bacterial communities.Results Compared with the dust exposed group,the top 5 bacterial phyla in the alveolar lavage fluid level of patients with pneumoconiosis were the same,followed by Proteobacteria,Firmicutes,Bacteroidetes,Fusobacteria,and Actinobacteria.Compared with the dust exposure group,the pneumoconiosis group patients belong to the top 5 genera of horizontal flora abundance,which are different.The dust exposure group is respectively:Pseudomonas,Proctor,Streptococcus,Achromobacter,and Neisseria.The pneumoconiosis group is respectively:Pseudomonas,Achromobacter,Streptococcus,Ralstonia,and Proctor.The Alpha diversity analysis results showed that compared with the dust exposed group,the level of bacterial diversity in the pneumoconiosis group was difference(P<0.05),and there was no statistically significant difference in bacterial evenness(P>0.05);Beta diversity showed differences in microbial community structure between the two groups(P<0.05).Single factor microbial association network analysis showed that there was a high correlation between Firmicutes and Bacteroidetes in the pneumoconiosis and dust exposed groups and other species,showing a positive correlation;The correlation between Proteobacteria and other species is high,showing a negative correlation.Conclusion The structure and relative abundance of bacteria in lower respiratory tract were different between patients with pneumoconiosis and dust exposure,and the diversity of bacteria in lower respiratory tract increased in patients with pneumoconiosis,which may be related to disease status.