高通量测序探究流产女性及未孕女性子宫内的菌群
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摘要
目的·探究流产女性及未孕女性的子宫中是否存在微生物菌群。方法·选取复旦大学附属华东医院妇科通过负压吸引刮宫术获得的56例自然流产女性和39例人工流产女性的子宫蜕膜组织,以及通过宫腔镜手术获得的10例未孕女性(排除感染)的子宫内膜组织。选取16S核糖体RNA(rRNA)基因高可变区V4对3组样本进行高通量测序,分析各组样本的微生物组组成,并比较各组间菌群多样性的差异。结果·在流产女性的蜕膜组织及未孕女性的子宫内膜组织中均检测到了低丰度的微生物组。3组样本中,均以变形菌门(Proteobacteria)为主要优势菌门,大肠埃希菌/志贺菌属(Escherichia/Shigella)为主要优势菌属。自然流产女性蜕膜组织的菌群多样性高于人工流产女性。结论·子宫内存在以变形杆菌门(Proteobacteria)为优势菌的低丰度菌群。自然流产女性与人工流产女性子宫菌群中的优势菌在门、科、属水平上无显著差异,但自然流产女性子宫菌群多样性高于人工流产女性。
Objective · To explore the uterine microbiota in women undergoing abortion and non-pregnant women. Methods · Fifty-six women who had experienced spontaneous abortion and 39 women who had experienced artificial abortion were selected to obtain the decidual tissues by curettage vacuum aspiration, and 10 non-pregnant women(without infection) were selected to obtain the endometrial tissues through gynecological surgery from Department of Gynecology in Huadong Hospital. Three groups of samples were subjected to high-throughput sequencing based on the V4 variable region of the 16 S ribosomal RNA(rRNA) gene, and the bacterial composition of each group of samples was analyzed. The diversity of uterine microbiota was compared among three groups. Results · Low-abundance microbiomes were detected from the decidual tissues of abortion women and the endometrial tissues of non-pregnant women. Proteobacteria was the main phylum of the three groups, and Escherichia/Shigella was the main genus. The diversity of uterine microbiota in spontaneous abortion group was higher than that in artificial abortion group. Conclusion · There is a low-abundance uterine microbiota mainly composed of Proteobacteria. The dominant bacteria at the level of phylum, family and genus has no significant difference between the spontaneous abortion group and the artificial abortion group, but there is higher flora diversity in the spontaneous abortion group than in the artificial abortion group.
Objective · To explore the uterine microbiota in women undergoing abortion and non-pregnant women. Methods · Fifty-six women who had experienced spontaneous abortion and 39 women who had experienced artificial abortion were selected to obtain the decidual tissues by curettage vacuum aspiration, and 10 non-pregnant women(without infection) were selected to obtain the endometrial tissues through gynecological surgery from Department of Gynecology in Huadong Hospital. Three groups of samples were subjected to high-throughput sequencing based on the V4 variable region of the 16 S ribosomal RNA(rRNA) gene, and the bacterial composition of each group of samples was analyzed. The diversity of uterine microbiota was compared among three groups. Results · Low-abundance microbiomes were detected from the decidual tissues of abortion women and the endometrial tissues of non-pregnant women. Proteobacteria was the main phylum of the three groups, and Escherichia/Shigella was the main genus. The diversity of uterine microbiota in spontaneous abortion group was higher than that in artificial abortion group. Conclusion · There is a low-abundance uterine microbiota mainly composed of Proteobacteria. The dominant bacteria at the level of phylum, family and genus has no significant difference between the spontaneous abortion group and the artificial abortion group, but there is higher flora diversity in the spontaneous abortion group than in the artificial abortion group.
引文
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[27]Zhu L,Luo F,Hu W,et al.Bacterial communities in the womb during healthy pregnancy[J].Front Microbiol,2018,9:2163.
[28]Collado MC,Rautava S,Aakko J,et al.Human gut colonisation may be initiated in utero by distinct microbial communities in the placenta and amniotic fluid[J].Sci Rep,2016,6:23129.
[29]Antony AK,Aagaard K,Ganu RS,et al.The placenta harbors a unique microbiome[J].Sci Transl Med,2014,6(237):237ra65.
[2]Hillier SL,Nugent RP,Eschenbach DA,et al.Association between bacterial vaginosis and preterm delivery of a low-birth-weight infant.The Vaginal Infections and Prematurity Study Group[J].N Engl J Med,1995,333(26):1737-1742.
[3]Taylor DP,Darville T,Haggerty CL.Does bacterial vaginosis cause pelvic inflammatory disease?[J].Sex Transm Dis,2013,40(2):117-122.
[4]Hillier SL,Kiviat NB,Hawes SE,et al.Role of bacterial vaginosis-associated microorganisms in endometritis[J].Am J Obstet Gynecol,1996,175(2):435-441.
[5]Nelson DB,Bellamy S,Nachamkin I,et al.First trimester bacterial vaginosis,individual microorganism levels,and risk of second trimester pregnancy loss among urban women[J].Fertil Steril,2007,88(5):1396-1403.
[6]Heinonen PK,Teisala K,Punnonen R,et al.Anatomic sites of upper genital tract infection[J].Obstet Gynecol,1985,66(3):384-390.
[7]Eschenbach DA,Rosene K,Tompkins LS,et al.Endometrial cultures obtained by a triple-lumen method from afebrile and febrile postpartum women[J].JInfect Dis,1986,153(6):1038-1045.
[8]Hemsell DL,Obregon VL,Heard MC,et al.Endometrial bacteria in asymptomatic,nonpregnant women[J].J Reprod Med,1989,34(11):872-874.
[9]M?ller BR,Kristiansen FV,Thorsen P,et al.Sterility of the uterine cavity[J].Acta Obstet Gyn Scan,2011,74(3):394-396.
[10]Mitchell CM,Haick A,Nkwopara E,et al.Colonization of the upper genital tract by vaginal bacterial species in nonpregnant women[J].Am J Obstet Gynecol,2015,212:611.e1-611.e9.
[11]Perez-MunOz ME,Arrieta MC,Ramer-Tait AE,et al.A critical assessment of the“sterile womb”and“in utero colonization”hypotheses:implications for research on the pioneer infant microbiome[J].Microbiome,2017,5(1):48.
[12]Zervomanolakis I,Ott HW,Hadziomerovic D,et al.Physiology of upward transport in the human female genital tract[J].Ann N Y Acad Sci,2007,1101:1-20.
[13]Hansen LK,Becher N,Bastholm S,et al.The cervical mucus plug inhibits,but does not block,the passage of ascending bacteria from the vagina during pregnancy[J].Acta Obstet Gynecol Scand,2013,93(1):102-108.
[14]Kunz G,Beil D,Deiniger H,et al.The uterine peristaltic pump.Normal and impeded sperm transport within the female genital tract[J].Adv Exp Med Biol,1997,424:266-267.
[15]Moreno I,Codo?er FM,Vilella F,et al.Evidence that the endometrial microbiota has an effect on implantation success or failure[J].Am J Obstet Gynecol,2016,215(6):684-703.
[16]Fang RL,Chen LX,Shu WS,et al.Barcoded sequencing reveals diverse intrauterine microbiomes in patients suffering with endometrial polyps[J].Am JTransl Res,2016,8(3):1581-1592.
[17]Franasiak J,Werner M,Juneau C,et al.Endometrial microbiome at the time of embryo transfer:next-generation sequencing of the 16S ribosomal subunit[J].JAssist Reprod Gen,2016,33(1):129-136.
[18]Khan KN,Fujishita A,Masumoto H,et al.Molecular detection of intrauterine microbial colonization in women with endometriosis[J].Eur J Obstet Gynecol Reprod Biol,2016,199:69-75.
[19]Moreno I,Franasiak JM.Endometrial microbiota:new player in town[J].Fertil Steril,2017,108:32-39.
[20]Verstraelen H,Vilchezvargas R,Desimpel F,et al.Characterisation of the human uterine microbiome in non-pregnant women through deep sequencing of the V1-2 region of the 16S rRNA gene[J].Peerj,2016,4(1):e1602.
[21]Walther-António MR,Chen J,Multinu F,et al.Potential contribution of the uterine microbiome in the development of endometrial cancer[J].Genome Med,2016,8(1):122.
[22]Chen C,Song X,Wei W,et al.The microbiota continuum along the female reproductive tract and its relation to uterine-related diseases[J].Nat Commun,2017,8(1):875.
[23]Miles SM,Hardy BL,Merrell DS.Investigation of the microbiota of the reproductive tract in women undergoing a total hysterectomy and bilateral salpingooopherectomy[J].Fertil Steril,2017,107:813-820.
[24]Tao X,Franasiak JM,Zhan Y,et al.Characterizing the endometrial microbiome by analyzing the ultra-low bacteria from embryo transfer catheter tips in IVFcycles:next generation sequencing(NGS)analysis of the 16S ribosomal gene[J].Hum Microbiome J,2017,3:15-21.
[25]Ravel J,Gajer P,Abdo Z,et al.Vaginal microbiome of reproductive-age women[J].Proc Natl Acad Sci U S A,2011,108(Suppl 1):4680-4687.
[26]Witkin SS,Linhares IM.Why do lactobacilli dominate the human vaginal microbiota?[J].BJOG,2017,124(4):606-611.
[27]Zhu L,Luo F,Hu W,et al.Bacterial communities in the womb during healthy pregnancy[J].Front Microbiol,2018,9:2163.
[28]Collado MC,Rautava S,Aakko J,et al.Human gut colonisation may be initiated in utero by distinct microbial communities in the placenta and amniotic fluid[J].Sci Rep,2016,6:23129.
[29]Antony AK,Aagaard K,Ganu RS,et al.The placenta harbors a unique microbiome[J].Sci Transl Med,2014,6(237):237ra65.