Diversity And Difference Of Oral Flora In Patients With Ankylosing Spondylitis Of Two TCM Syndromes

Objective:To investigate the relationship of oral flora disorder in ankylosing spondylitis patients with TCM syndrome by analyzing the differences of oral flora among ankylosing spondylitis patients with kidney deficiency and wetness-heat syndrome,kidney deficiency and du meridian cold syndrome and healthy controls..And to study the correlation between oral flora disorder and immune disorders in ankylosing spondylitis disease in order to find a new directions for the pathogenesis of ankylosing spondylitis and clinical treatment.Methods: 33 ankylosing spondylitis patients with kidney deficiency and du meridian cold syndrome,33 ankylosing spondylitis patients with kidney deficiency and wetness-heat syndrome and 31 healthy controls were enrolled in the study.The case report form was used to collect the patient’s clinical information of Chinese and Western medicine.The non-irritating saliva of all subjects were collected to extract DNA for16 Sr RNA high-throughput sequencing using Illumina Miseq platform.The oral flora composition of all subjects was detected and analyzed to look for the differences between ankylosing spondylitis patients and healthy controls,and between ankylosing spondylitis patients with kidney deficiency and du meridian cold syndrome and patients with kidney deficiency and wetness-heat syndrome.Cytokines in plasma from all subjects including IL-10,IL-18 and SIg A and SIg A in saliva were detected by enzyme-linked immunosorbent assay(ELISA)competition method to analyze oral flora abundance and cytokines levels for finding the correlation between the oral flora and mucosal immunity in ankylosing spondylitis patients.Result: 1.Results found that it consisted of 17 Phylums,32 classes,63 orders,115 families,and 224 Genus in the oral cavity of all subjects in our study.The results from ɑdiversity analysis showed that the abundance and diversity of oral flora in ankylosing spondylitis patients group were greater than those in healthy group(P<0.05).Although there was a certain difference of the composition of the oral flora between ankylosing spondylitis patients group and the healthy group,there was no statistic difference in βdiversity(P>0.05).Compared with healthy control group,The difference analysis showed that there were 1 species in the Phylums level,3 species in the family level,11 species in the genus level of bacteria different between AS patients and healthy volunteers(p<0.05).2.The levels of IL-10,IL-18 and SIg A in plasma of patients with ankylosing spondylitis were lower than those in healthy group(P<0.05).The level of SIg A in saliva was not different from that in healthy group(P>0.05).Correlation analysis showed that there are 29 species are associated with ESR or CRP in patients,19 species are associated with ASDAS or BASDAI,11 species are associated with IL-10 or IL-18 or SIg A,and 26 species are associated with saliva SIg A in the genus level,(P<0.05).The Fusobacteriaceae_unclassified in the oral cavity of AS patients was positively correlated with ESR and ASDAS-ESR,while Leptotrichiaceae-other was positively correlated with IL-10 in plasma,and the abundance of two colonies in AS oral cavity was higher than that in normal subjects.3.The ɑ diversity and β diversity analysis of patients with ankylosing spondylitis showed that the abundance and composition of the oral flora of the two syndromes were not significantly different(P>0.05),There were some difference in the abundance of oral flora between two type of syndromes including 2 species in the phylums level、3 species in the family level and 7 species in the genus level(P<0.05)。Conclusions:1.There is a disorder in the oral flora of patients with ankylosing spondylitis.2.Mucosal immune function may be inhibited in patients with ankylosing spondylitis,and the disorder of oral flora were associated with disease activity and mucosal immune abnormalities.3.There is no significant difference in the overall abundance and diversity of oral flora between patients with the syndrome of Kidney deficiency and wetness-heat and the syndrome of kidney deficiency and du meridian cold.However,there are differences in the abundance of several floras at the genus level.The link between TCM symptoms of ankylosing spondylitis and oral flora needs further investigation.