Correlation between Changes of Vaginal Microflora and Premature Rupture of Membranes during Pregnancy
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摘要:
目的 通过对正常妊娠与胎膜早破孕妇的阴道微生物群落分析，阐明阴道菌群变化与胎膜早破的相关性。 方法 随机收集符合纳入标准的不同孕周阶段胎膜早破孕妇阴道分泌物50例为实验组，正常孕妇阴道分泌物30例为正常对照组，提取细菌DNA后用高通量测序技术，分析比较胎膜早破与正常孕妇阴道微生物群落的多样性指数及群落结构。 结果 各组的Alpha多样性丰富度及均匀度指数，差异无统计学意义（P > 0.05），各组菌群非度量多维尺度分析（nonmetric multidimensional scaling, NMDS）Stress = 0.02,差异具有统计学意义（ P < 0.05）。根据维恩图显示未足月胎膜早破组的物种多样性最高，为529个；未足月正常对照组组的多样性最低，为207个。 结论 正常孕期阴道菌群较稳定，Alpha多样性小,Beta多样性显著。妊娠期阴道微生物群落以卷曲乳杆菌为主，若孕期阴道内菌群多样性增加，惰性乳杆菌、加德纳菌、纤毛菌、普氏菌、奇异菌等异常增加，易发生细菌性阴道病、胎膜早破。 Abstract:Objective To elucidate the relationship between vaginal microbiota and premature rupture of membranes by analyzing the vaginal microbiota of normal pregnancy and pregnant women of premature rupture of membranes. Methods Gestational age meeting inclusion criteria was randomly collected （gestational age < 28 weeks; 28 to 34 weeks; 34 and 37 weeks; 37 to 39 weeks; each case was 10 at 39-41 weeks） , a total of 50 cases of vaginal secretions from pregnant women with premature rupture of membranes were used as the experimental group. Vaginal secretions were collected from 6 normal pregnant women at the above relative gestational age, and a total of 30 normal women were selected as the control group. The bacterial DNA of vaginal secretions was extracted by the second generation high throughput sequencing method, and the diversity index, microbial community structure and dominant species of vaginal microbiome were analyzed and compared between premature rupture of membranes and normal pregnant women. Results The P value of Wilcoxon test for Alpha diversity richness and evenness index among each group was > 0.05, The Stress value of nonmetric multidimensional scaling was = 0.02, less than 0.05, the difference was statistically significant. According to Venn diagram, the species diversity of preterm premature rupture of membranes was the highest （529）. The diversity of normal control group before term was the lowest （207）. Conclusions In normal pregnancy, vaginal microflora is stable, with small Alpha diversity and significant Beta diversity. Vaginal microbial community during pregnancy can be divided into five types, mainly L. crispatus , if the increase of L.iners, gardnella, prevotella,sneathia , ureaplasma and atopobium etc in the vagina during pregnancy, bacterial vaginosis and premature rupture of membranes are prone to occur. -
图 2 足月/未足月胎膜早破组与足月/未足月对照组稀释曲线
Figure 2. Rarefaction curve of PPROM，WZY，PROM and ZY group
图 4 未足月/足月胎膜早破组与未足月/足月对照组的微生物群落非度量多维尺度分析
Figure 4. The NMDS of PPROM，WZY，PROM and ZY group
图 5 胎膜早破与对照组在门水平等级微生物物种组成
Figure 5. Microbial species composition at phylum level of TMZPZ and control group
图 6 胎膜早破组与对照组在属水平等级微生物物种组成
Figure 6. Microbial species composition at genus level of TMZPZ and Control
图 7 样本在种水平等级上的微生物群落类型聚类图
注释：柱状图中不同颜色代表不同的物种相对丰度，根据柱状图、树状聚类图可主要分为以下几种微生物群落类型（community state types，CST），红色：CST I，蓝色：CST III ，黄色：CST IV ， 深蓝色：CST V ，绿色：CST II 。
Figure 7. Cluster diagram of microbial community types at species level
图 8 样本微生物群落在种水平分布维恩图
Figure 8. Venn diagram of sample microbial community distribution at species level
图 9 胎膜早破组与对照组的Circos图
注释：左半部分是丰度top5的物种以及其对应的丰度信息，右半部分是按照分组展示的分组信息，所占宽度越宽，表示丰度越高。
Figure 9. Circos diagram of TMZPZ and control group
表 1 各分组丰富度及均匀度指数的多样性分析
Table 1. Diversity analysis of the richness and evenness indices of each grouping
组别 shannon simpson chao1 对照组 1.01 ± 0.26 0.34 ± 0.09 67.46 ± 14.31 TMZPZ组 1.02 ± 0.36 0.35 ± 0.13 68.85 ± 23.5 t 0.058 0.390 0.328 P 0.954 0.697 0.744 WTY组 1.06 ± 0.29 0.36 ± 0.1 69 ± 14.97 ZY组 0.94 ± 0.18 0.31 ± 0.07 65.16 ± 13.55 PPROM组 1.07 ± 0.39 0.38 ± 0.15 70.04 ± 18.36 足月PROM组 0.94 ± 0.3 0.31 ± 0.09 67.05 ± 30.07 F 0.973 1.566 0.193 P 0.410 0.205 0.901 
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