Objective

2

To investigate the difference of mouth flora between

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

Methods

2

Twenty female with pSS were included and divided into

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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

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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 （

P

＞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

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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 （

P

＜0.01）. Totally， 2

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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

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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

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deficiency and fluid depletion group （

P

＜ 0.05 or

P

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

P

＞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

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

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deficiency and fluid depletion group decreased significantly （

P

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

P

＞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

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deficiency and fluid depletion group （

P

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

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

P

＜0.05 or

P

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

P

＞0.05）.

Conclusion

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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

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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.