胎膜早破孕妇胎盘与胎膜菌群丰度的高通量测序研究  

High-throughput sequencing study on the abundance of placenta and fetal membrane flora in pregnant women with premature rupture of membranes

作　　者：何霞 黄蓉 祁文瑾[1] Xia He;Rong Huang;Wenjin Qi(Department of Obstetrics,the First Affiliated Hospital of Kunming Medical University,Kunming 650032,Yunnan Province,China)

机构地区：[1]昆明医科大学第一附属医院产科,昆明650032

出　　处：《中华妇幼临床医学杂志（电子版）》2024年第5期549-555,共7页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：国家自然科学基金项目(82260302);云南省医疗卫生单位内设研究机构科研项目(2018NS0141)。

摘　　要：目的探讨胎膜早破(PROM)孕妇及正常孕妇胎盘与胎膜菌群丰度的高通量测序结果及其差异。方法选择2019年1月至2020年2月于昆明医科大学第一附属医院产科分娩的15例孕妇为研究对象。根据其是否发生PROM及其发生PROM时孕龄,将其分别纳入未足月PROM(PPROM)组(n=5,PPROM孕妇),足月PROM(TPROM)组(n=6,TPROM孕妇)与对照组(n=4,未发生PROM孕妇)。3组孕妇均于胎盘娩出后,立即采集胎儿面与母体面胎盘及胎膜组织样本各1份,提取胎盘、胎膜组织中菌群DNA,对其16S核糖体RNA(rRNA)基因的高可变区(V3、V4区)进行高通量测序,并对3组孕妇胎盘、胎膜组织样本中菌群类别、构成进行分析。采用Metastats软件,对PPROM组与对照组、TPROM组与对照组、PPROM组与TPROM组胎盘与胎膜组织细菌菌属相对丰度进行非参数T检验。3组孕妇分娩时年龄及体重等一般临床资料比较,差异均无统计学意义(P>0.05)。本研究遵循的程序符合2013年修订的《世界医学协会赫尔辛基宣言》要求。结果①3组孕妇分娩孕龄比较,差异有统计学意义(F=18.65,P<0.001)。②门水平,3组孕妇胎儿面与母体面胎盘及胎膜组织样本中相对丰度>0.1%的菌门,均为7种:厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、螺旋菌门(Spirochaetes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、蓝藻菌门(Cyanobacteria)、梭杆菌(Fusobacteria)。③属水平,3组孕妇胎儿面与母体面胎盘及胎膜组织样本中相对丰度>0.1%的菌属,均为6种:未分类菌属(uncultured bacterium)、乳酸杆菌属(Lactobacillus)、梭菌属(Clostridium)、链球菌属(Streptococcus)、链杆菌属(Catenibacterium)、密螺旋体属(Treponema)。④对3组孕妇胎儿面与母体面胎盘及胎膜组织样本属水平菌群差异性分析结果显示,PPROM组、TPROM组孕妇胎儿面与母体面胎盘及胎膜组织样本中梭菌属、链球菌属、未分类菌属、密螺旋体属相对丰度,均显�Objective To investigate the results of high-throughput sequencing of placenta and membrane flora abundance in premature rupture of membranes(PROM)and normal pregnant women,and to analyze the differences.Methods Fifteen pregnant women who gave birth in the Department of Obstetrics,the First Affiliated Hospital of Kunming Medical University from January 2019 to February 2020 were selected as study subjects.According to whether PROM occurred or not and their gestational age at the time of PROM occurrence,they were included into preterm PROM(PPROM)group(n=5,PPROM pregnant women),term PROM(TPROM)group(n=6,TPROM pregnant women)and control group(n=4,pregnant women without PROM).Immediately after placenta delivery,one sample of tissue from fetal face and maternal face placenta and fetal membrane was collected from pregnant women in three groups.The flora DNA in placenta and fetal membrane tissues was extracted,and the high variable region V3-V4 of 16S ribosomal RNA(r RNA)gene was sequenced with high throughput,and the flora structure of phylum and genus in the placenta and fetal membrane tissue samples of pregnant women in three groups was analyzed.Metastats software was used to compare the relative abundance of bacteria in placenta and fetal membranes tissues in PPROM group and control group,TPROM group and control group,PPROM group and TPROM group by parameter T test.There were no significant differences in the general clinical data such as age and weight of pregnant women during delivery among three groups(P>0.05).The procedure followed in this study met the requirements of the WorldMedicalAssociationDeclaration ofHelsinki revised in 2013.Results①The difference in gestational age of pregnant women among three groups was statistically significant(F=18.65,P<0.001).②At the phylum level,there were 7 phyla with relative abundance>0.1%in the placental and fetal membrane tissue samples of pregnant women in three groups:Firmicutes,Bacteroidetes,Spirochaetes,Proteobacteria,Actinobacteria,Cyanobacteria,and Fusobacteria.③

关 键 词：胎膜早破 高通量核苷酸序列分析 RNA 核糖体 16S 胎盘 细菌 生物多样性 菌群丰度 孕妇 

分 类 号：R714[医药卫生—妇产科学]