蔗糖刺激前后无龋人群和高龋患者唾液微生物群落差异分析

Analysis of difference of saliva microbiome between caries-free and caries-active individuals before and after sucrose treatment

目的研究蔗糖刺激前后无龋人群与高龋患者唾液微生物群落组成差异,探索造成差异的潜在关键微生物。方法本研究纳入无龋人群(CF组)8名,高龋患者(CA组)10例,分别于处理前和蔗糖溶液处理30 min、2 h后采集唾液样本,利用Illumina测序平台分析比较微生物多样性和群落结构。结果蔗糖处理前,CF组和CA组物种多样性和丰富度无统计学差异,处理2 h后,CA组微生物多样性显著高于CF组(P<0.05);Beta多样性分析结果显示两组未见明显差异。两组唾液菌群优势物种种类相同,以假单胞菌属、链球菌属、普雷沃菌属、拟普雷沃菌属、奈瑟菌属及韦荣菌属为主要优势菌,但细菌丰度不同。与处理前相比,蔗糖处理后两组韦荣菌在处理前后组间差异变化不明显,普雷沃菌和拟普雷沃菌差异呈增大趋势,奈瑟菌差异呈减小趋势。LEfSe分析显示,蔗糖刺激后,优势菌之一普雷沃菌在CA组的丰度显著高于CF组,提示其作为生物标志物的潜力。结论蔗糖刺激可导致高龋患者唾液菌群多样性增加,唾液微生物组成改变;龋病情况影响唾液微生物对蔗糖的反应;普雷沃菌具有作为潜在龋病生物标志物的作用。

Objective To investigate,before and after sucrose challenge,the difference of saliva microbiome between caries-free(CF)and caries-active(CA)individuals,and to explore the potential key microorganisms that is more susceptible to sucrose challenge.Methods Eighteen systemic healthy volunteers,among which 8 were caries-free and 10 were caries-active,were recruited.Saliva samples were collected pre-treatment and at 30 min and 2 h after the sucrose treatment.Microbial diversity and community structure were analyzed and compared using Illumina sequencing platform.Results The diversity and abundance of saliva samples from caries-free individuals was similar to that from caries-active individuals before sucrose treatment.After 2 h of treatment,the microbial diversity of the CA group was significantly higher than that of the CF group(P<0.05).The results of Beta diversity analysis showed no significant difference between the two groups.The dominant species,which were represented as Pseudomonas,Streptococcus,Prevotella,Alloprevotella,Neisseria and Veillonella,in the two groups were similar but different in abundance.There was no significant change in the abundance difference of Veillonella between CF and CA before and after treatment.The abundance differences of Prevotella and Alloprevotella between CF and CA increased and the abundance difference of Neisseria decreased after sucrose treatment.The results of LEfSe analysis showed that Prevotella,as one of the dominant bacteria in the two groups,was significantly more abundant in the CA group than in the CF group after sucrose treatment,suggesting its potential biomarker role.Conclusion The diversity and composition of saliva microbiome changes subject to sucrose challenge,which is more significant in caries-affected individuals.The response of salivary microbe to sucrose challenge is different between caries-free and caries-active individuals.Prevotella is a potential biomarker of caries.