基于16sRNA高通量测序技术分析粉尘接触与煤矿工人咽部菌群变化关联性

Analysis on the association between dust exposure and pharyngeal microbiota in coal miners based on 16sRNA high-throughput sequencing technology

目的探讨煤矿粉尘接触人群咽部菌群分布情况。方法采用判断抽样法,选择从事职业性粉尘接触工作20年以上的8名煤矿工人为接尘组,以工作中不接触粉尘的4名煤矿工人为对照组。采用咽拭子采集2组人群的咽拭子,提取咽部菌群,采用16sRNA基因高通量测序技术进行基因测序,分析菌群的多样性、菌群丰度和均匀度。结果对照组人群的咽部菌群组成的操作分类单元有254个,较接尘组多210个。与对照组比较,接尘组人群咽部菌落的Chao1指数、Shannon指数、PD-tree指数和Pielou指数均降低(P值均<0.01)。在门水平上,接尘组人群咽部拟杆菌门和梭杆菌门丰度均高于对照组(P值均<0.05);在属水平上,与对照组比较,接尘组人群咽部普氏菌属、奈瑟菌属和单胞菌属等丰度均增高(P值均<0.05),乳杆菌属丰度降低(P值<0.05)。受试者工作特征曲线分析结果显示,乳杆菌属、梭杆菌属和罗氏菌属对接尘工人咽部菌群失衡有较高的预测价值,曲线下面积均为1.00±0.00。结论煤矿粉尘接触人群咽部菌群物种多样性和菌群均匀度均降低,可能与煤尘的持续吸入扰乱了人体咽部的微生物环境有关。

Objective To explore the distribution of pharyngeal microbiota in coal miners exposed to dust.Methods Eight coal miners who had been engaged in occupational dust exposure for more than 20 years were selected as the dust-exposed group,and four coal miners who were not exposed to dust at work were selected as the control group using the judgment sampling method.Pharyngeal secretions of the coal miners were collected with throat swabs,and its pharyngeal microbiota was analyzed.The diversity,abundance and evenness of the microbiota were analyzed by gene sequencing using the 16sRNA gene highthroughput sequencing technology.Results A total of 254 operational taxonomic units of pharyngeal microbiota were detected in the coal miners in the control group,which was 210 more than that in the dust-exposed group.The Chao1 index,Shannon index,PD-tree index and Pielou index of pharyngeal microbiota in the dust-exposed group decreased compared with the control group(all P<0.01).The abundance of Bacteroidetes and Clostridum,at the phylum level,in the pharynx of coal miners in the dust-exposed group was higher than that in the control group(all P<0.05).The abundance of Prevotella,Neisseria,and Monas,at the genus level,in the pharynx of coal miners in the dust-exposed group was higher than that in the control group(all P<0.05),while the abundance of Lactobacillus decreased(P<0.05).The analysis results of the receiver operating characteristic curve showed that Lactobacillus,Fusobacterium and Rothia may play a role for pharyngeal microbiota imbalance prediction in dust-exposed workers,and the area under the curves were all 1.00±0.00.Conclusion The species diversity and evenness of pharyngeal microbiota in coal miners exposed to dust are decreased,which may be related to the continuous inhalation of coal dust that disrupts the microbial environment of the throat.