口源性口臭患者口腔不同生态位菌群及代谢分析_黄志强.pdf

基金项目:新疆维吾尔自治区高层次人才引进项目(人社明电 13 111 号)作者简介:黄志强，男，1996 08 生，硕士，E-mail:kimhuangzhiqiang163 com收稿日期:2022 11 30口源性口臭患者口腔不同生态位菌群及代谢分析黄志强，程永波*(新疆医科大学第一附属医院消化病一科，乌鲁木齐830054;*通讯作者，E-mail:chengyb2001 163com)摘要:目的研究口源性口臭患者与健康受试者口腔不同生态位菌群组成和代谢差异，探讨口源性口臭与口腔菌群的关系。方法随机选择 7 例以口臭为主诉就诊的口源性口臭患者(halitosis 组)以及 3 例健康受试者(normal 组)。分别记录两组口气值，收集两组唾液、舌苔、颊黏膜和颚黏膜菌群样本。采用 16S rNA 方法对样本进行物种组成和多样性分析，并采用 PICUSt 方法对样本进行代谢预测分析，组间差异显著性检验采用 Adonis 分析。结果两组 4 个生态位菌群丰度和多样性均无明显差异(均 P0 05)。Adonis 分析表明，两组间唾液菌群差异显著(P=0 011)，两组间舌苔(P=0 25)、颊黏膜(P=0 128)和颚黏膜(P=0 178)菌群结构相似。门水平上，halitosis 组唾液中螺旋体门、颊黏膜中螺旋体门和拟杆菌门、颚黏膜中放线菌门比例均明显升高(均 P0 05)。属水平上，halitosis 组弯曲杆菌属和巨球型菌属在唾液、舌苔和颊黏膜中均表现出高比例，密螺旋体属在唾液和颊黏膜中比例明显升高，纤毛菌属在舌苔和颊黏膜中比例升高(均 P 0 05)。此外，halitosis组唾液中小杆菌属、舌苔中棒状杆菌属、颊黏膜中普雷沃菌属和毛绒厌氧杆菌属也表现出高比例(均 P0 05)，而颚黏膜中仅巨球型菌属比例增高(P=0 017)。代谢方面，两组口腔 4 个生态位在硫及含硫氨基酸代谢未表现出显著差异(均 P 0 05)，但 halitosis 组唾液中泛酸盐和 CoA 生物合成途径明显增强，颊黏膜中糖胺聚糖降解途径及烟酸和烟酰胺代谢途径明显增强，颚黏膜中糖酵解/糖异生途径明显增强(均 P0 05)。结论口腔不同生态位致臭菌的高丰度定植、微生物失衡和代谢变化与口源性口臭的发生密切相关。关键词:口源性口臭;口腔菌群;菌群失衡;16S rNA 基因测序;代谢中图分类号:78文献标志码:A文章编号:1007 6611(2023)04 0498 09DOI:1013753/j issn1007 6611202304013Microbial and metabolic analysis in different ecological sites of oral cavity of patients with intra-oral halitosisHUANG Zhiqiang，CHENG Yongbo*(First Department of Gastroenterology，First Affiliated Hospital of Xinjiang Medical University，Urumqi 830054，China;*Corresponding author，E-mail:chengyb2001163 com)Abstract:ObjectiveTo investigate the correlation between intra-oral halitosis and oral flora by studying the difference in the compo-sition and metabolism of different ecological sites of oral flora between patients with intra-oral halitosis and healthy subjectsMethodsSeven patients who presented with a complaint of halitosis(halitosis group)and three healthy subjects(normal group)were randomlyenrolled to record the breath values and collect the samples of saliva，coating on the tongue，buccal mucosa，and jaw mucosa floraThe samples were analyzed for species composition and diversity using the 16S rNA gene sequencing technology and the metabolicprediction analysis was conducted using the PICUSt method，and the difference between groups was tested using Adonis analysisesultsThere was no significant difference in the abundance and the diversity of the four ecological sites between two groups(all P0 05)Adonis analysis showed that except salivary microbiota(P=0 011)，the microbial structures in coating on the tongue(P=0 25)，buccal mucosa(P=0 128)and jaw mucosa(P=0 178)were similar between two groups At the phylum level，the propor-tions of Spirochaetes in the saliva，Spirochaetes and Bacteroidetes in the buccal mucosa，and Actinobacteria in the jaw mucosa weresignificantly higher in halitosis group than in normal group(all P 0 05)At the genus level，the proportions of Campylobacter andMegasphaera were higher in saliva，coating on the tongue，and buccal mucosa in halitosis group than in normal group，the proportionsof Treponem in saliva and buccal mucosa，and Leptotrichia in coating on the tongue and buccal mucosa were significantly higher in hali-tosis group than in normal group(all P0 05)In addition，the proportions of Dialister in saliva，Corynebacterium in the coating onthe tongue，Prevotella and Lachnoanaerobaculum in the buccal mucosa were significantly higher in halitosis group than in normal group(all P0 05)，and only the proportion of Megasphaera in the jaw mucosa was higher in halitosis group(P=0 017)The sulfur andsulfur-containing amino acid metabolism at the four ecological sites in the oral cavity showed no significant difference between two groups(all P005)，but the pantothenate and CoA biosynthesis pathways were obviously enhanced in the saliva in halitosis group，the glyco-saminoglycan degradation pathway and the nicotinic acid and nicotinamide metabolism pathway were significantly enhanced in the894J Shanxi Med Univ，Apr 2023，Vol 54 No 4buccal mucosa，and the glycolysis/gluconeogenesis pathway were significantly enhanced in the jaw mucosa in halitosis group(all P 0 05)ConclusionHigh abundance colonization of odor-causing bacteria，microbial imbalance and metabolic changes at differentecological sites of the oral cavity are closely associated with the development of intra-oral halitosisKey words:intra-oral halitosis;oral microbiota;microbiota imbalance;16S rNA gene sequencing;metabolism口源性口臭(intra-oral halitosis，IOH)是一种常见的口腔菌群失衡性疾病，长期反复的症状会使患者产生沟通恐惧、焦虑抑郁、自卑苦恼等不利情绪，严重影响人们的社会交往和身心健康1 3。近年来4 7，通过高通量测序技术发现具核梭杆菌(Fu-sobacterium nucleatum)、牙龈卟啉单胞菌(Porphy-romonas gingivalis)、齿密垢螺旋体(Treponema denti-cola)、摩尔梭菌(Solobacterium moorei)、中间普雷沃菌(Prevotella intermedia)、福赛斯坦纳菌(Tannerellaforsythia)、消化链球菌(Peptostreptococcus)以及纳米细菌等都与 IOH 密切相关，并通过体外培养技术证实了它们可分解含硫底物(半胱氨酸、蛋氨酸、色氨酸、精氨酸和赖氨酸等含硫氨基酸8 10)，进而释放挥发性硫化物(volatile sulfur compounds，VSCs)产生臭味。但口腔中约有 500 700 种细菌11，12，其基因库多样性大约是宿主的 100 倍13，因此针对产臭菌的研究仍在继续。此外，随着代谢组学的发展，越来越多的研究者也将焦点集中在菌群的复杂代谢功能，而不是其组成。因口腔处于开放的环境，有关IOH 患者口腔不同生态位菌群结构和差异的研究相对较少且有较大分歧7，14，15。本研究以 16S rNA基因测序技术为手段，通过检测 IOH 患者与健康受试者唾液、舌苔、颊黏膜和颚黏膜样本中微生物组成，分析其口腔菌群结构差异，并通过 PICUSt 代谢预测方