首段肺灌洗液是16S rRNA测序法分析尘肺病患者肺部菌群信息的更优选择  

The first-stage lung lavage solution is a better choice for analyzing the lung microbiota information of patients with pneumoconiosis by 16S rRNA sequencing

作　　者：邓冠华 唐侍豪 肖芹 张海[1] 邝嘉惠 王致[1] DENG Guanhua;TANG Shihao;XIAO Qin;ZHANG Hai;KUANG Jiahui;WANG Zhi(Key Laboratory of Occupational Environment and Health,Guangzhou Twelfth People's Hospital,Guangdong Guangzhou,510620,China;School of Basic Medicine and Public Health,Jinan University,Guangzhou Guangdong,510632,China;First Internal Medicine Department,Guangzhou Haizhu District Hospital of Traditional Chinese Medicine,Guangzhou Guangdong,510230,China)

机构地区：[1]广州市第十二人民医院职业环境与健康重点实验室,广东广州510620 [2]暨南大学基础医学与公共卫生学院,广东广州510632 [3]广州市海珠区中医医院内一科,广东广州510230

出　　处：《职业与健康》2023年第18期2461-2465,2473,共6页Occupation and Health

基　　金：广州市卫生健康科技项目(20201A010036);广州市科学技术局重点研发计划项目(202206010061);广州市医学重点学科建设项目(2021—2023年);广州市卫生健康科技重大项目(2021A031003)。

摘　　要：目的探索基于高通量测序评估首段肺灌洗液对研究尘肺病患者肺部菌群多样性的意义。方法2021年采集不同尘肺病患者或不同疾病场景各时段肺灌洗液34份,采用十六烷基三甲基溴化铵法提取样本总基因组DNA,扩增16S rRNA V4区域,对经Illumina NovaSeq测序得到的样本序列进行拼接、标准化质控,并进行生物信息学分析。结果各样本序列质控有效率达77.13%。较中后段肺灌洗液,首段肺灌洗液门水平物种以厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)和放线菌门(Actinobacteria)为主,拟杆菌门未被低估,抗炎治疗前后也未被低估。首段和各时段肺灌洗液的物种数和Shannon指数比较,差异均无统计学意义(均P>0.05),各段肺灌洗液的肺部菌群丰富度和均一性特征相似。不同个体首段肺灌洗液物种数和Shannon指数比较,差异均有统计学意义(均P<0.05),首段肺灌洗液可区分不同个体肺部菌群特征。抗炎治疗前后首段肺灌洗液物种数比较,差异有统计学意义(P<0.05);Shannon指数比较,差异无统计学意义(P>0.05),首段肺灌洗液可分辨抗炎场景下肺部菌群特征变化。首段和各时段肺灌洗液Beta多样性比较,差异无统计学意义(P>0.05),各段肺灌洗液的肺部菌群组成相似。个体间和抗炎治疗前后首段肺灌洗液Beta多样性比较,差异均有统计学意义(均P<0.05),首段肺灌洗液可分辨不同干预场景肺部菌群组成的变化,也可反映肺部菌群组成的个体性差异。结论首段肺灌洗液更灵敏反映尘肺病患者肺部菌群组成信息,是高通量16S rRNA标签测序法分析职业性肺病菌群信息的更优选择。Objective To explore the significance of evaluating first-stage lung lavage fluid based on high-throughput sequencing in the study of pulmonary bacterial diversity in pneumoconiosis patients.Methods A total of 34 lung lavage fluid samples were collected in different pneumoconiosis patients or different disease scenarios at different time periods.Genomic DNA was extracted from each sample according to the sixteen alkyl three methyl bromide(CTAB)instructions.The V4 primers of 16S rRNA gene was amplified.The sample sequences obtained by Illumina NovaSeq sequencing were spliced,standardized quality control,and bioinformatics analysis was performed.Results Effective tags of samples was 77.13%.Compared with the middle and posterior lung lavage fluid,the horizontal species of the first lung lavage fluid group were dominated by Firmicutes,Proteobacteria,Bacteroidota and Actinobacteria,and the Bacteroidete was not underestimated,nor before and after antibiotic treatment.There were no difference in the richness and Shannon index of lung lavage fluid collected in the first segment and other time period(both P>0.05).Flora richnessand evenness in all segments of lung lavage fluid were similar.There were significant differences of the richness and Shannon index of lung lavage fluid in the first segment of lung lavage fluid in different individuals(both P<0.05).The first segment lung lavage fluid can distinguish the characteristics of lung flora in different individuals.There was significant difference in the richness of first segment lung lavage fluid before and after antibacterial treatment(P<0.05),but no difference in Shannon index(P>0.05).The first segment lung lavage fluid can distinguish the changes of pulmonary flora characteristics in the anti-inflammatory scenario.The Beta diversity was no difference in between the first and other segment lung lavag(P>0.05),but significant differences were found between individuals and before and after anti-inflammatory treatment(both P<0.05).The first segment lung lavage fluid can dis

关 键 词：尘肺 微生物组 16S rRNA 灌洗液 

分 类 号：R135.2[医药卫生—劳动卫生]