O Dorien F, Roskams Tania, Van den Eynde Kathleen, Vanhie Arne, Peterse Daniëlle P, Meuleman Christel, Tomassetti Carla, Peeraer Karen, D'Hooghe Thomas M, Fassbender Amelie
1 Department of Development and Regeneration, Organ systems, KU Leuven, Leuven, Belgium.
2 Department of Obstetrics and Gynaecology, Leuven University Fertility Centre, University Hospital Leuven, Leuven, Belgium.
Reprod Sci. 2017 Feb;24(2):242-251. doi: 10.1177/1933719116653677. Epub 2016 Sep 27.
To reinforce Sampson's theory of retrograde menstruation in the pathogenesis of endometriosis, proof should be provided that during menstruation endometrial cells are present in peritoneal fluid (PF). We hypothesize that the prevalence of PF samples containing endometrial cells is higher in patients with endometriosis than in controls without endometriosis during menstruation. We selected from our biobank PF samples of 17 reproductive-age women with (n = 9) or without (n = 8) endometriosis who had received a diagnostic laparoscopy for investigation of pain/infertility. Peritoneal fluid had been collected during laparoscopy in the menstrual phase of the cycle, centrifuged, and the resulting pellet was stored at -80°C. About 5-μm sections of frozen PF pellets were stained using the Dako Envision Flex system with primary antibodies against epithelial cell adhesion molecule (Ep-CAM; endometrial epithelial cells), CD10 (endometrial stromal cells), prekeratin (epithelial/mesothelial cells), vimentin (endometrial/mesothelial/immune cells), calretinin (mesothelial cells), and CD68 (macrophages). The PF cells positive for Ep-CAM were detected in 5 of 9 patients with endometriosis and 6 of 8 controls ( P = .62). CD10 stained positively in 6 of the 9 patients with endometriosis and 3 of the 8 controls ( P = .35). Calretinin and prekeratin staining showed the presence of mesothelial cells in all pellets. Vimentin stained approximately 100% of the PF cells. CD68+ macrophages represented >50% of cells in all pellets. The prevalence of PF samples containing endometrial epithelial and stromal cells was not higher in patients with endometriosis than in controls without endometriosis during menstruation. Our findings question the relevance of endometrial cells in PF for the pathogenesis of endometriosis and support the importance of other mechanisms such as immune dysfunction and/or endometrial stem cells.
为了强化桑普森关于逆行月经在子宫内膜异位症发病机制中的理论，应提供证据证明在月经期间腹膜液（PF）中存在子宫内膜细胞。我们假设，在月经期间，患有子宫内膜异位症的患者中含有子宫内膜细胞的PF样本的患病率高于无子宫内膜异位症的对照组。我们从生物样本库中选取了17名接受诊断性腹腔镜检查以调查疼痛/不孕的育龄妇女的PF样本，其中9名患有子宫内膜异位症，8名没有（对照组）。在月经周期的月经期腹腔镜检查期间收集腹膜液，离心后，所得沉淀保存在-80°C。使用Dako Envision Flex系统对冷冻PF沉淀的约5μm切片进行染色，使用针对上皮细胞粘附分子（Ep-CAM；子宫内膜上皮细胞）、CD10（子宫内膜间质细胞）、角蛋白前体（上皮/间皮细胞）、波形蛋白（子宫内膜/间皮/免疫细胞）、钙视网膜蛋白（间皮细胞）和CD68（巨噬细胞）的一抗。在9名子宫内膜异位症患者中的5名和8名对照组中的6名中检测到Ep-CAM阳性的PF细胞（P = 0.62）。9名子宫内膜异位症患者中的6名和8名对照组中的3名CD10染色呈阳性（P = 0.35）。钙视网膜蛋白和角蛋白前体染色显示所有沉淀中均存在间皮细胞。波形蛋白染色约100%的PF细胞。CD68+巨噬细胞在所有沉淀中占细胞的比例>50%。在月经期间，患有子宫内膜异位症的患者中含有子宫内膜上皮和间质细胞的PF样本的患病率并不高于无子宫内膜异位症的对照组。我们的研究结果质疑了PF中的子宫内膜细胞与子宫内膜异位症发病机制的相关性，并支持了免疫功能障碍和/或子宫内膜干细胞等其他机制的重要性。
