早期类风湿关节炎女性患者阴道微生态菌群基因组分析

目的 从菌群总体角度分析和比较早期类风湿关节炎患者和健康育龄期女性阴道菌群差异.方法 采集女性新发未治早期类风湿关节炎患者(RA组) 及健康育龄期女性(对照组) 各12例的阴道分泌物,提取样本细菌总基因组DNA,然后进行16S rRNA V3-V4区基因扩增及采用Illumina高通量测序技术对扩增的PCR产物进行测序,再采用Uparse、LEfSe等软件,分析两组样本阴道菌群物种丰度和阴道菌群结构.结果 RA组阴道菌群Alpha多样性显著增加,差异有统计学意义(P = 0.04) .健康对照组阴道菌群以乳酸杆菌为优势菌属,占84.98％.RA组患者阴道菌群中乳酸杆菌属则显著减少(11.01％) ,加德纳菌属显著增加(43.16％) ,同时双歧杆菌属(9.94％) 、链球菌属(8.05％) 、奇异菌属(7.86％) 等相对丰度亦增加.PCoA主坐标分析和样本层级聚类分析的结果 表明: RA组阴道菌群结构有较高相似性,与健康对照组阴道菌群结构可明显区分.LEfSe分析结果 显示,在属水平上,两组乳杆菌属、加德纳菌属、双歧杆菌属、奇异菌属等丰度差异有统计学意义(P值为0.017、0.046、0.018、0.007) .结论 阴道菌群失调与类风湿关节炎之间存在相关性,RA患者阴道菌群多样性增加,乳杆菌显著减少,加德纳菌等厌氧菌属相对丰度增高.

Genomic analysis of vaginal microbiome community in female early rheumatoid arthritis patients

Objective To analyze the differences of vaginal microbiome between the patients with early rheumatoid arthritis (RA) and healthy reproductive-age women from the overall perspective of bacteria. Methods A total of 24 vaginal swabs were collected from 12 early RA patients and 12 healthy women (control). The bacterial genomic DNA of all obtained samples was extracted for amplifying the 16S rRNA V3and V4 hypervariable region by PCR and pyrosequencing by Illumina. The abundance and composition of vaginal microbiota were analyzed by Uparse and LEfSe software. Results The Alpha diversity was significantly increased in the bacterial strains from the early RA patients (P = 0.04). Lactobacillus was the predominant genus in healthy women (accounting for 84.98％). The amount of Lactobacillus was decreased(11.01％) but that of Gardnerella increased (43.16％) significantly in the RA group. Moreover, the relative abundances of Bifidobacterium (9.94％), Streptococcus (8.05％) and Atopobium (7.86％) were increased in different degrees. The results of principal coordinate analysis (PCoA) and hierarchical cluster analysis showed that the structure of vaginal flora in RA group had high similarity, and the vaginal microflora structure of the healthy control group could be distinctly distinguished. LEfSe analysis found that there were significant differences in the abundance of Lactobacillus, Gardiner, bifidobacteria and Singularities at the genus level (P = 0.017, 0.046, 0.018, 0.007). Conclusion A correlation is found between vaginal dysbacteriosis and RA. The diversity of vaginal microbiome is increased in the RA patients, with significantly decreased relative abundance of Lactobacillus, while increased abundance of Gardnerella.