Khan Khaleque N, Fujishita Akira, Masumoto Hiroshi, Muto Hideki, Kitajima Michio, Masuzaki Hideaki, Kitawaki Jo
Department of Obstetrics and Gynecology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan; Department of Obstetrics and Gynecology, Graduate School of Biomedical Sciences, Nagasaki University, Japan.
Saiseikai Nagasaki Hospital, Nagasaki, Japan.
Eur J Obstet Gynecol Reprod Biol. 2016 Apr;199:69-75. doi: 10.1016/j.ejogrb.2016.01.040. Epub 2016 Feb 6.
Increased intrauterine microbial colonization by bacteria culture method and occurrence of endometritis have been reported in women with endometriosis. Here we investigated microbial colonization in intrauterine environment and cystic fluid of women with and without endometriosis by molecular approach.
This is a case-controlled biological study with a total of 32 women each with and without endometriosis. Among them, 16 each in these two groups of women received treatment with gonadotropin-releasing hormone agonist (GnRHa). Pattern of microbial colonization in endometrial swabs and endometrioma/non-endometrioma cystic fluid was examined using broad-range polymerase-chain reaction (PCR) amplification of bacteria targeting 16S rRNA gene (rDNA). After quantification of index PCR product, 16S rDNA metagenome sequence analysis was done by Illumina Miseq system.
A wide proportion (0.01-97.8%) of multiple bacteria was detected in both endometrial swabs and cystic fluid collected from women with and without endometriosis. 16S metagenome assay indicated that proportion of Lactobacillacae was significantly decreased (p<0.01) and of Streptococcaceae, Staphylococaceae, Enterobacteriaceae was significantly increased (p<0.05 for each) in GnRHa-treated women with endometriosis than in GnRHa-untreated women. While bacteria culture method failed to detect a single colony, 16S metagenome assay could detect significantly higher percentage of Streptococcaceae (p<0.01) and Staphylococaceae (p<0.05) in the cystic fluid derived from women with ovarian endometrioma comparing to that in cystic fluid collected from non-endometrioma cysts.
These findings indicate the occurrence of sub-clinical infection in intrauterine environment and in the cystic fluid of ovarian endometrioma. Additional side effect of GnRHa treatment in promoting silent intrauterine and/or ovarian infection should be considered.
据报道，采用细菌培养方法发现子宫内膜异位症女性的子宫内微生物定植增加，且出现了子宫内膜炎。在此，我们通过分子方法研究了有或无子宫内膜异位症女性的子宫内环境和囊液中的微生物定植情况。
这是一项病例对照生物学研究，共有32名患有和未患有子宫内膜异位症的女性。其中，两组女性各有16名接受了促性腺激素释放激素激动剂（GnRHa）治疗。使用针对16S rRNA基因（rDNA）的细菌广谱聚合酶链反应（PCR）扩增，检测子宫内膜拭子和子宫内膜瘤/非子宫内膜瘤囊液中的微生物定植模式。在对指数PCR产物进行定量后，通过Illumina Miseq系统进行16S rDNA宏基因组序列分析。
在患有和未患有子宫内膜异位症的女性收集的子宫内膜拭子和囊液中均检测到多种细菌的广泛比例（0.01 - 97.8%）。16S宏基因组分析表明，与未接受GnRHa治疗的女性相比，接受GnRHa治疗的子宫内膜异位症女性中乳酸杆菌科的比例显著降低（p<0.01），而链球菌科、葡萄球菌科、肠杆菌科的比例显著增加（各p<0.05）。虽然细菌培养方法未能检测到单个菌落，但与从非子宫内膜瘤囊肿收集的囊液相比，16S宏基因组分析在来自卵巢子宫内膜瘤女性的囊液中能检测到显著更高比例的链球菌科（p<0.01）和葡萄球菌科（p<0.05）。
这些发现表明子宫内环境和卵巢子宫内膜瘤囊液中存在亚临床感染。应考虑GnRHa治疗在促进隐匿性子宫内和/或卵巢感染方面的额外副作用。
