复发性流产患者阴道微生物宏基因组分析

Metagenomic analysis of vaginal flora in patients with recurrent spontaneous abortion

摘要:

目的：研究和分析复发性流产患者阴道菌群微生物多样性差异、种类差异及功能分析，初步阐释阴道微生态在复发性流产发生发展过程中起到的作用。方法：选取2010年1月至2016年12月纳入中山大学第八附属医院妇产科的复发性流产患者21例，正常妊娠人工流产20例作为本课题的研究对象，2组在怀孕时间上不存在统计学差异（P>0.05）；同时收集整理各种临床诊疗数据。术前常规进行病史采集并取阴道分泌物，利用宏基因组测序分析微生物类型及微生物基因组数据，明确差异性及功能性。结果：临床数据显示复发性流产患者阴道炎症发生率高于正常妊娠人群；复发性流产患者阴道微生物多样性明显高于正常妊娠人群，微生物基因丰度低于正常妊娠人群。通过差异性物种鉴定分析，挑选得到上百种差异性微生物，结合样本实际情况及统计学要求，选取16种差异性微生物，其中5种菌株在正常妊娠人群阴道中没有发现，而在复发性流产患者阴道中明显存在且丰度较高；8种菌株在正常妊娠人群中存在且丰度较高，但在复发性流产患者阴道中没有检测到；3种菌株丰度差异较大。这些差异性微生物的功能富集分析显示差异性菌群在分子功能、生物过程、细胞组成存在统计学差异，具体体现在氨基酸代谢、糖代谢、能量代谢、核苷酸代谢、脂代谢等代谢过程，尤其是各种氨基酸代谢与能量代谢。此外，本研究还发现代谢功能异常的影响还会引起阴道内部细胞信号转导异常，引起内分泌与代谢疾病心血管疾病的风险。结论：复发性流产患者免疫低下可能与阴道微生态紊乱引起的阴道炎症有关，除不同种的乳酸菌互换外，加德纳菌、普氏菌等新菌种可能也在复发性流产的发生发展过程中起到了重要作用。

Abstract:

Objective：To study and analyze the differences in microbial diversity and species differences of vaginal flora caused by recurrent spontaneous abortion，and to preliminarily explain the role of vaginal microecology in the development of recurrent sponta－neous abortion. Methods：21 patients with recurrent spontaneous abortion admitted to our hospital from January 2010 to December 2016 and 20 cases of induced abortion to normal pregnancy were selected as subjects in this study. There was no significant difference in pregnancy time between the two groups（P>0.05），and various data for clinical diagnosis and treatment were collected at the same time. After the routine collection of medical history，vaginal secretions were collected. Metagenomic sequencing was used to analyze microbial types and microbial genomic data of the secretions，so as to clarify differences and functionality. Results：Clinical data showed that the incidence of vaginal inflammation in patients with recurrent spontaneous abortion was higher than that in normal pregnancy. The vaginal microbial diversity of patients with recurrent spontaneous abortion was significantly higher than that of normal pregnancy，and the microbial gene abundance was lower than that of normal pregnancy. Through differential species identification analysis，hundreds of different differential microorganisms were selected，and 16 different micro-organisms were selected according to statistical requirements and the actual conditions of the sam－ples. Five of the strains were not found in the vagina of normal pregnant people，but clearly present in the vagina of patients with recurrent spontaneous abortion and had a high abun－dance；8 strains were present in the vagina of normal pregnant people and had a high abundance，but were not detected in the vagina of patients with recurrent spontaneous abortion；the other three strains had large differences in abundance. The functional en－richment analysis of these differential microorganisms showed that there are statistical differences in molecular function，biological processes and cell composition among them，which were reflected in metabolic processes such as amino acid metabolism，glucose metabolism，energy metabolism，nucleotide metabolism and lipid metabolism，and various amino acid metabolism and energy metabolism in particular. In addition，we also found that the influence of metabolic dysfunction can also cause abnormal vaginal in－ternal cell signal transduction，endocrine and metabolic disease，causing cardiovascular disease in endocrine and metabolic diseases in return. Conclusion：The low immunity of patients with recurrent spontaneous abortion may be associated with vaginal inflammation caused by vaginal microecological disorders. In addition to the exchange of different types of lactic acid bacteria，new strains such as Gardnerella and Plasmodium may also play an important role in the occurrence and development of recurrent spontaneous abortion.