基于16S rRNA的肝郁血虚型失眠患者口腔微生态与中医舌象的相关性研究

Study on the Correlation between Oral Microecology and Tongue Coating of Insomnia Patients with Liver Depression and Blood Deficiency based on 16S rRNA

目的探究肝郁血虚型失眠患者不同中医舌象口腔菌群的物种组成与生物多样性的异同。方法临床收集肝郁血虚型失眠受试者口腔菌群样本29例,其中淡白舌舌象13例,紫暗舌舌象16例。对样本进行16S rRNA测序、数据处理,并使用生物信息学与统计学方法对比分析菌群的生物多样性及物种组成。结果物种注释及组成分析表明,肝郁血虚型失眠患者主要由15个菌门中的245个菌属构成。门水平上,5种优势菌门相对丰度共占比96.48%:厚壁菌门(43.13%)、变形菌门(20.60%)、放线菌门(11.07%)、拟杆菌门(13.78%)、梭杆菌门(7.9%)。属水平上,7种优势菌属相对丰度共占比68.32%:链球菌(31.14%)、奈瑟菌(10.21%)、罗斯氏菌(7.62%)、梭杆菌(4.06%)嗜血杆菌(3.73%)。淡白舌组α生物多样性显著高于紫暗舌组。Anosim统计分析,两组组间R值为0.169,p值为0.006,表明两组口腔菌群组间差异具有显著性。LDAsize分析表明,两组在20种微生物菌群方面存在显著性差异,可作为鉴别生物标志物进一步分析,包括罗斯氏菌属、奈瑟菌属、梭杆菌属等。结论肝郁血虚型失眠患者淡白舌组与紫暗舌组的口腔菌群在生物多样性与物种组成方面均具有显著的差异性,可为进一步探究肝郁血虚型失眠口腔微生态特征及阐释淡白舌与紫暗舌的差异性机理提供借鉴。

Objective To explore the similarities and differences of species composition and biodiversity of oral flora of different tongue signs in insomnia patients with liver depression and blood deficiency.Methods 29 oral microflora samples of insomnia patients with liver depression and blood deficiency were collected,including 13 pale tongue and 16 purple and dark tongue.The samples were sequenced and processed by 16S rRNA,and the biodiversity and species composition of bacteria were analyzed by bioinformatics and statistics.Results At the phylum level,the relative abundance of five dominant phylums was 96.48%:Firmicutes(43.13%),Proteobacteria(20.60%),Actinobacteria(11.07%),Bacteroidetes(13.78%)and Fusobacteria(7.9%).At generic level,the relative abundance of seven dominant genera was 68.32%:Streptococcus(31.14%),Neisseria(10.21%),Rothia(7.62%),Fusobactericum(4.06%)and Haemophilus(3.73%).The biodiversity of alpha in light white tongue group was significantly higher than that in purple dark tongue group.Anosim statistical analysis showed that the R value was 0.169 and P value was 0.006 between the two groups,indicating that there was significant difference between the two groups.Species annotation and composition analysis showed that all samples consisted of 245 genera of 15 phyla.LDA size analysis showed that there were significant differences in 20 microbial flora between the two groups,which could be used as a biomarker for further analysis,including Rothia,Neisseria and Fusobactericum.Conclusion There are significant differences in biodiversity and species composition between light white tongue group and purple dark tongue group in patients with insomnia due to liver depression and blood deficiency,which can provide reference for further exploring the oral microecological characteristics of insomnia due to liver depression and blood deficiency and explaining the differential mechanism between light white tongue and purple dark tongue.