基于16S rRNA测序分析肾虚证先兆流产的阴道菌群特征

Structural Characterization of Vaginal Microbiota in Threatened Abortion Patients with Syndrome of Kidney Deficiency by 16S rRNA Sequencing

目的:探讨肾虚证早期先兆流产的阴道菌群结构特征。方法:选取2018年5月至2020年12月就诊于广州中医药大学第一附属医院的肾虚证早期先兆流产患者(DK-TA组)31例、正常早孕女性(NP组)116例,收集两组人群的阴道分泌物,采用16S rRNA测序技术,对两组阴道菌群群落组成及差异菌进行分析。结果:与NP组比较,Alpha多样性显示DK-TA组阴道菌群丰富度及多样性显著增高。通过Binary jaccard及unweighted_unifrac两种距离矩阵分析可知两组阴道菌群的相似性、分散性、丰度及进化关系存在明显差异。在门水平上,与NP组比较,DK-TA组的拟杆菌门、梭杆菌门相对丰度降低,而变形菌门及芽单胞菌门相对丰度明显增高;在属水平上,与NP组比较,DK-TA组纤毛菌属、双歧杆菌属相对丰度降低,而志贺氏菌属、卫星沙特尔沃思氏菌属明显升高。线性判别分析(LEfSe)拟杆菌门、梭杆菌门、双歧杆菌属为NP组阴道优势菌种。变形菌门、厚壁菌门是DK-TA组的阴道优势菌种。功能预测发现DK-TA组与环腺苷3’,5’-单磷酸信号通路、炎症介质对色氨酸通道的调节等38个功能路径密切相关,且两组阴道差异菌与差异代谢路径存在明显的正负关联。结论:变形菌门、芽单孢菌门、拟杆菌门可能是表征肾虚证早期先兆流产的阴道菌群生物标志物。

Objective:To investigate the structural features of vaginal microbiota in the early threatened abortion patients with the syndrome of kidney deficiency.Method:Thirty-one patients with early threatened abortion of kidney deficiency syndrome(DK-TA group)and 116 women with normal early pregnancy(NP group)attending the First Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2018 to December 2020 were selected.The vaginal secretions were collected for 16S rRNA sequencing,which can reveal the vaginal microbiota composition and differential bacteria between the two groups.Result:The DK-TA group had higher abundance and diversity of vaginal microbiota than the NP group.The Binary jaccard and unweighted_unifrac distance matrix analysis showed that the similarity,dispersion,abundance,and phylogenetic relationship of vaginal microbiota were significantly different between the two groups.At the phylum level,the DK-TA group had lower relative abundance of Bacteroidetes and Fusobacteria and higher relative abundance of Proteobacteria and Gemmatimonadetes than the NP group.At the genus level,the DK-TA group had lower relative abundance of Sneathia and Bifidobacterium and higher relative abundance of Escherichia-Shigella and Shuttleworthia than the NP group.Linear discriminant analysis Effect Size(LEfSe)revealed that Bacteroidetes,Fusobacteria,and Bifidobacteria were dominant in the NP group and Proteobacteria and Firmicutes in the DK-TA group.The function prediction found that DK-TA was closely associated with 38 functional pathways,including cyclic adenosine monophosphate signaling pathway and regulation of tryptophan channels by inflammatory mediators.In addition,the vaginal differential bacteria between the two groups had significant positive or negative association with the differential metabolic pathways.Conclusion:Proteobacteria,Gemmatimonadetes,and Bacteroidetes in the vaginal microbiota may be biomarkers for threatened abortion of kidney deficiency syndrome.