Characteristics of Mouth Flora in 20 Primary Sjogren's Syndrome Patients with Different Patterns

BAI Zhiyi; LI Yuan; WANG Haiyu; LU Siyi; BAI Xuejiao; ZHU Yuelan; HOU Xiujuan

doi:10.13288/j.11-2166/r.2023.01.009

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Characteristics of Mouth Flora in 20 Primary Sjogren's Syndrome Patients with Different Patterns

更新时间：2022-12-29

- Characteristics of Mouth Flora in 20 Primary Sjogren's Syndrome Patients with Different Patterns
- Journal of Traditional Chinese Medicine Vol. 64, Issue 1, Pages: 43-48(2023)
- 作者机构：
  
  1.北京中医药大学，北京市朝阳区北三环东路11号，100029
  2.北京中医药大学东方医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2023.01.009
  CLC：
- Received：26 May 2022，
  
  Revised：2022-09-09，
  
  Published：02 January 2023
- 稿件说明：
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柏志义,李媛,王海瑜等.20例不同证型原发性干燥综合征患者口腔菌群特征[J].中医杂志,2023,64(01):43-48.

BAI Zhiyi,LI Yuan,WANG Haiyu,et al.Characteristics of Mouth Flora in 20 Primary Sjogren's Syndrome Patients with Different Patterns[J].Journal of Traditional Chinese Medicine,2023,64(01):43-48.
柏志义,李媛,王海瑜等.20例不同证型原发性干燥综合征患者口腔菌群特征[J].中医杂志,2023,64(01):43-48. DOI： 10.13288/j.11-2166/r.2023.01.009.

BAI Zhiyi,LI Yuan,WANG Haiyu,et al.Characteristics of Mouth Flora in 20 Primary Sjogren's Syndrome Patients with Different Patterns[J].Journal of Traditional Chinese Medicine,2023,64(01):43-48. DOI： 10.13288/j.11-2166/r.2023.01.009.

摘要

目的

观察并比较阴虚津亏型与津液失布型原发性干燥综合征（pSS）患者口腔菌群的差异。

方法

20例pSS女性患者辨证分为阴虚津亏组9例，津液失布组11例，另将8例健康女性设为正常组。收集各组受试者年龄及pSS患者病程等一般资料，采用口干视觉模拟评分（VAS）及简化版口腔干燥量表（SXI）评估患者口干严重程度；收取所有受试者口腔含漱液，抽提口腔含漱液样本细菌总DNA进行PCR扩增，针对16S rDNA基因的V3-V4区进行Illumina NovaSeq高通量测序，得到口腔菌群生物信息数据。基于有效数据进行组间可操作分类单元（OTU）分析；采用线性判别分析法分析组间群落差异；采用Chao1指数、Shannon指数、observed OTU指数分析物种 Alpha多样性，并进行组间Beta多样性分析。

结果

两组患者病程及各组受试者年龄比较差异均无统计学意义（

＞0.05）。津液失布组口干VAS评分为（4.00±1.73）分，阴虚津亏组为（7.00±2.06）分；津液失布组SXI评分为（12.45±1.81）分，阴虚津亏组为（20.33±3.54）分，两组VAS评分、SXI评分差异均有统计学意义（

＜0.01）。口腔菌群测序后获得有效序列2 366 361条，聚类后正常组、津液失布组、阴虚津亏组分别获得1288、1989、1770个特征OTU。与正常组比较，津液失布组、阴虚津亏组厚壁菌门丰度均升高，阴虚津亏组梭杆菌门、卟啉单胞菌属丰度降低，韦荣氏球菌属丰度升高（

＜0.05或

＜0.01）；津液失布组与阴虚津亏组各菌门、各菌属水平比较差异均无统计学意义（

＞0.05）。正常组优势菌门为梭杆菌门、螺旋体门、软壁菌门、互养菌门，优势菌属为梭杆菌属、密螺旋体属、卟啉单胞菌属、不动杆菌属；津液失布组优势菌门为GN02、SR1，优势菌属为链球菌属；阴虚津亏组优势菌门为厚壁菌门，优势菌属为韦荣氏球菌属、放线菌属。与正常组比较，阴虚津亏组Shannon指数明显下降（

＜0.05），Chao1指数、observed OTU指数差异无统计学意义（

＞0.05）；与正常组及阴虚津亏组比较，津液失布组各指数差异均无统计学意义（

＞0.05）。Beta多样性分析显示，正常组与津液失布组、阴虚津亏组口腔菌群组成结构差异均具有统计学意义（

＜0.05或

＜0.01），阴虚津亏组与津液失布组组成结构差异无统计学意义（

＞0.05）。

结论

不同证型的pSS患者口腔微环境各有其特征性优势菌，阴虚津亏型患者口腔菌群组成结构变化及优势菌丰度变化均较津液失布型患者更显著。

Abstract

Objective

To investigate the difference of mouth flora between

yin

deficiency and fluid depletion pattern and failure of body fluids distribution pattern in people with primary Sjogren's syndrome （pSS）.

Methods

Twenty female with pSS were included and divided into

yin

deficiency and fluid depletion group （9 cases） and failure of body fluids distribution group （11 cases）， and eight healthy women were set as normal group. General information including age， disease course were collected， and the severity of dry mouth was evaluated by visual analogue scale （VAS） and summated xerostomia inventory （SXI）. Oral gargle samples were collected， and total bacterial DNA was extracted for PCR amplification. Illumina NovaSeq high-throughput sequencing was performed for V3-V4 region of 16S rDNA gene to obtain bioinformation data of mouth flora. Intergroup operational taxonomic unit （OTU） analysis was performed based on valid data， and linear discriminant analysis was used to analyze community difference between groups. The Chao1 index， Shannon index， and observed OTU index were used to analyze the alpha diversity of species， and the Beta diversity analysis between groups was carried out.

Results

There was no statistical difference in the course of disease between groups of different patterns， as well as in the age among all the three groups （

＞0.05）. The VAS score of dry mouth in the failure of body fluids distribution group was 4.00±1.73， while that in the

yin

deficiency and fluid depletion group was 7.00±2.06； correspondingly， the SXI score of the two groups was 12.45±1.81 and 20.33±3.54； there were significant differences in the VAS score and SCI score between the two groups （

＜0.01）. Totally， 2

366

361 effective sequences were obtained after sequencing， and 1288 characteristic OTU were obtained through clustering in the normal group， 1989 in the failure of body fluids distribution group and 1770 in the

yin

deficiency and fluid depletion group， respectively. Compared to those in normal group， the abundance of Firmicutes significantly increased in the other two groups， and the abundance of Fusobacterium and Porphyromonas decreased， while the abundance of Veillonella increased in the

yin

deficiency and fluid depletion group （

＜ 0.05 or

＜0.01）. There was no significant difference between the two groups with different TCM patterns in different bacterial phylum and genus levels（

＞0.05）. The dominant bacteria phylum in the normal group were Fusobacteria， Spirochaetes， Tenericutes and Synergistetes， while the dominant bacterial genera were Fusobacterium， Treponema， Porphyromonas， and Acinetobacter. GN02 and SR1 were the dominant bacterial phyla in the failure of body fluids distribution group， and Streptococcus was the dominant genera. Firmicutes were the dominant phyla in the

yin

deficiency and fluid depletion group， and the dominant genera were Veillonella and Actinomycetes. Compared to those in the normal group， Shannon index in the

yin

deficiency and fluid depletion group decreased significantly （

＜0.05）， while Chao1 index and observed OTU index showed no statistical difference （

＞0.05）. There was no statistical difference in Shannon index， Chao1 index and observed OTU index comparing the failure of body fluids distribution group to the normal group and the

yin

deficiency and fluid depletion group （

＞0.05）. Beta diversity analysis showed that there was statistically significant difference in the composition structure when comparing the normal group to the

yin

deficiency and fluid depletion group and the failure of body fluids distribution group （

＜0.05 or

＜0.01）， while there was no statistically significant difference between the two groups with different TCM patterns （

＞0.05）.

Conclusion

There are differences in the oral microenvironment of pSS with different TCM patterns， and each has its characteristic dominant bacteria； the changes of mouth flora composition and structure in

yin

deficiency and fluid depletion group are more significant than those in the failure of body fluids distribution group， as well as the changes of dominant bacteria abundance.

关键词

Keywords

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