复发性流产患者阴道菌群特征

doi:10.12280/gjszjk.20220143

摘要/Abstract

摘要：

目的：分析复发性流产（recurrent spontaneous abortion,RSA）患者阴道菌群特征。方法：采集2020年12月—2021年9月就诊于山西省妇幼保健院的20例不明原因RSA患者（RSA组）和20例正常早孕人工流产患者（对照组）的阴道分泌物,采用16S rRNA高通量测序技术分析2组患者阴道菌群的差异。结果：α多样性分析显示,2组Chao1指数、Shannon指数、Observed_OTU指数比较,差异无统计学意义（均P>0.05）。β多样性分析显示,2组阴道细菌群落结构显著不同（P<0.05）。分类学分析显示,在门、科、属水平,2组共同的优势菌为假单胞菌（变形菌门）,假单胞菌在RSA组中门、科、属水平的丰度分别为22.8%、21.9%、21.9%,在对照组中的丰度分别为17.9%、17.3%、17.3%,RSA组假单胞菌的丰度明显增加。结论：RSA患者存在阴道菌群失衡,特定菌属（如假单胞菌）可能与RSA发生、发展有关。

关键词: 流产,习惯性, 阴道, 高通量核苷酸序列分析, 阴道菌群, 高通量测序

Abstract:

Objective: To analyze the characteristics of vaginal flora in patients with recurrent spontaneous abortion (RSA). Methods: The vaginal secretions of 20 patients with unexplained RSA (RSA group) and 20 women with normal early pregnancy abortion (control group) from December 2020 to September 2021 were collected. The vaginal flora of vaginal secretions were analyzed by 16S rRNA high-throughput sequencing technology. Results: The α diversity analysis showed that there were no significant differences in the Chao1 index, Shannon index, and Observed_OTU index between the two groups (all P>0.05). The β diversity analysis showed the significant difference in the vaginal bacterial community structure between the two groups (P<0.05). Taxonomic analysis showed that the common dominant bacteria at the gate, family and genus levels were Pseudomonas (Proteobacteria) in the two groups, and that the abundance of Pseudomonas in the RSA group at the gate, family and genus levels was 22.8%, 21.9%, and 21.9%, while the abundance of Pseudomonas in the control group was 17.9%, 17.3% and 17.3%, respectively. The abundance of Pseudomonas in the RSA group was increased significantly than that in the control group. Conclusions: Patients with RSA have vaginal flora imbalance, and the specific bacteria (such as Pseudomonas) may be related with the occurrence and development of RSA.

Key words: Abortion,habitual, Vagina, High-throughput nucleotide sequencing, Vaginal flora, High throughput sequencing

程阳, 赵敏. 复发性流产患者阴道菌群特征[J]. 国际生殖健康/计划生育, 2022, 41(3): 184-188.

CHENG Yang, ZHAO Min. Characteristics of Vaginal Flora in Patients with Recurrent Spontaneous Abortion[J]. Journal of International Reproductive Health/Family Planning, 2022, 41(3): 184-188.

图/表 6

表1 2组患者一般情况比较

组别	n	年龄（岁）	BMI（kg/m²）	终止妊娠时天数（d）	孕次（次）	产次（次）	流产次数（次）
RSA组	20	31.00（28.00,33.75）	23.73±2.71	75.50（69.25,85.00）	3.00（2.00,4.00）	0.00（0.00,1.00）	3.00（2.00,3.00）
对照组	20	32.00（26.25,34.00）	22.57±2.09	51.00（47.00,53.00）	2.50（2.00,3.75）	1.00（0.00,1.00）	2.00（1.25,2.00）
t或Z		0.164	1.520	5.28	1.003	1.977	3.612
P		0.870	0.137	<0.001	0.316	0.048	<0.001

图1 各个样品在各分类水平上的序列注释程度柱形图注：横坐标是样品名,A代表RSA组,B代表对照组,纵坐标表示注释到该水平的序列数目占总注释数据的比例,柱状图自上而下的颜色顺序对应于右侧的图例颜色顺序。

图2 2组阴道菌群共有或特有物种韦恩图注：韦恩图显示的是不同分组间共有或特有的OTU数量,每个椭圆代表1个分组。紫色区域代表RSA组,粉色区域代表对照组,重叠区域代表2组共同的OTU。

图3 2组阴道菌群α多样性指数比较注：A为Chao1指数,B为Shannon指数,C为Observed_OTU指数。

图4 2组阴道菌群β多样性分析注：横坐标（Axis.1）表示第一主成分,百分比表示第一主成分对样品差异的贡献值;纵坐标（Axis.2）表示第二主成分,百分比表示第二主成分对样品差异的贡献值。同一个组的样品使用同一种颜色表示。

图5 2组样本在门、科、属水平相对分布柱形图（相对丰度前20的物种）注：X轴代表2组样品,Y轴代表相对丰度比,不同颜色代表不同的属,不同色块高度代表不同物种的相对丰度。

参考文献 16

[1]	Bender Atik R, Christiansen OB, Elson J, et al. ESHRE guideline: recurrent pregnancy loss[J]. Hum Reprod Open, 2018, 2018(2): hoy004. doi: 10.1093/hropen/hoy004. doi: 10.1093/hropen/hoy004
[2]	Christiansen OB, Larsen EC, Egerup P, et al. Intravenous immunoglobulin treatment for secondary recurrent miscarriage: a randomised, double-blind, placebo-controlled trial[J]. BJOG, 2015, 122(4):500-508. doi: 10.1111/1471-0528.13192. doi: 10.1111/1471-0528.13192 URL
[3]	Farr A, Kiss H, Hagmann M, et al. Role of Lactobacillus Species in the Intermediate Vaginal Flora in Early Pregnancy: A Retrospective Cohort Study[J]. PLoS One, 2015, 10(12):e0144181. doi: 10.1371/journal.pone.0144181. doi: 10.1371/journal.pone.0144181 URL
[4]	Sharma H, Tal R, Clark NA, et al. Microbiota and pelvic inflammatory disease[J]. Semin Reprod Med, 2014, 32(1):43-49. doi: 10.1055/s-0033-1361822. doi: 10.1055/s-0033-1361822 URL
[5]	Soper DE, Mayhall CG, Dalton HP. Risk factors for intraamniotic infection: a prospective epidemiologic study[J]. Am J Obstet Gynecol, 1989, 161(3):562-566; discussion 566-568. doi: 10.1016/0002-9378(89)90356-6. doi: 10.1016/0002-9378(89)90356-6
[6]	Romero R, Gómez R, Chaiworapongsa T, et al. The role of infection in preterm labour and delivery[J]. Paediatr Perinat Epidemiol, 2001, 15(Suppl 2):41-56. doi: 10.1046/j.1365-3016.2001.00007.x. doi: 10.1046/j.1365-3016.2001.00007.x. URL
[7]	黄汉陵, 李阳. 复发性流产患者阴道菌群变化及其与凋亡蛋白Bcl-2和Bax的相关性[J]. 中国微生态学杂志, 2022, 34(2):216-219. doi: 10.13381/j.cnki.cjm.202202017. doi: 10.13381/j.cnki.cjm.202202017
[8]	Koedooder R, Mackens S, Budding A, et al. Identification and evaluation of the microbiome in the female and male reproductive tracts[J]. Hum Reprod Update, 2019, 25(3):298-325. doi: 10.1093/humupd/dmy048. doi: 10.1093/humupd/dmy048 pmid: 30938752
[9]	Zhang F, Zhang T, Ma Y, et al. Alteration of vaginal microbiota in patients with unexplained recurrent miscarriage[J]. Exp Ther Med, 2019, 17(5):3307-3316. doi: 10.3892/etm.2019.7337. doi: 10.3892/etm.2019.7337 pmid: 30988706
[10]	Fan T, Zhong XM, Wei XC, et al. The alteration and potential relationship of vaginal microbiota and chemokines for unexplained recurrent spontaneous abortion[J]. Medicine (Baltimore), 2020, 99(51):e23558. doi: 10.1097/MD.0000000000023558. doi: 10.1097/MD.0000000000023558 URL
[11]	Palleroni NJ. Pseudomonas[M]. Atlanta, GA: American Cancer Society, 2015.
[12]	Moran AP, Prendergast MM, Appelmelk BJ. Molecular mimicry of host structures by bacterial lipopolysaccharides and its contribution to disease[J]. FEMS Immunol Med Microbiol, 1996, 16(2):105-115. doi: 10.1111/j.1574-695X.1996.tb00127.x. doi: 10.1111/j.1574-695X.1996.tb00127.x. URL
[13]	Hall-Stoodley L, Costerton JW, Stoodley P. Bacterial biofilms: from the natural environment to infectious diseases[J]. Nat Rev Microbiol, 2004, 2(2):95-108. doi: 10.1038/nrmicro821. doi: 10.1038/nrmicro821 pmid: 15040259
[14]	Mulcahy LR, Isabella VM, Lewis K. Pseudomonas aeruginosa biofilms in disease[J]. Microb Ecol, 2014, 68(1):1-12. doi: 10.1007/s00248-013-0297-x. doi: 10.1007/s00248-013-0297-x URL
[15]	Wong G, Antani JD, Lele PP, et al. Roadmap on emerging concepts in the physical biology of bacterial biofilms: from surface sensing to community formation[J]. Phys Biol, 2021, 18(5):10.1088/1478-3975/abdc0e. doi: 10.1088/1478-3975/abdc0e. doi: 10.1088/1478-3975/abdc0e
[16]	Zhao F, Chen Y, Gao J, et al. Characterization of Vaginal Microbiota in Women With Recurrent Spontaneous Abortion That Can Be Modified by Drug Treatment[J]. Front Cell Infect Microbiol, 2021, 11:680643. doi: 10.3389/fcimb.2021.680643. doi: 10.3389/fcimb.2021.680643 URL