Ji Sifan, Li Yamei, Liang Guiling, Zhang Huiyu, Yan Li, Zhao Xiaoya, Chen Luting, Zhang Jian
Department of Obstetrics and Gynaecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, China.
Department of Obstetrics and Gynaecology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China; Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, China.
Reprod Biomed Online. 2025 Jun;50(6):104499. doi: 10.1016/j.rbmo.2024.104499. Epub 2024 Oct 22.
How does tubal endometriosis (TEM) impact the morphology and ultrastructure of tubal epithelium, and what are the underlying pathophysiological mechanisms of TEM?
Epithelial samples of fallopian tube were collected from patients with and without TEM (TEM group, n = 27; control group, n=28). Morphological characteristics, ultrastructure, percentage of ciliated cells and ciliary beat frequency (CBF) were assessed via haematoxylin and eosin staining, electron microscopy and high-speed camera. mRNA microarray analysis was conducted to identify differentially expressed genes (DEG) in tubal epithelium, which were further validated using quantitative polymerase chain reaction (qPCR), immunohistochemistry and Western blotting. The effects of recombinant chemokine ligand 2 (CCL2) protein on primary human fallopian tube epithelial cells (FTEC) were examined in vitro.
Patients with TEM exhibited abnormalities in the morphology and ultrastructure of tubal epithelium, with a significantly reduced percentage of ciliated cells (P < 0.0001) and CBF (P < 0.0001) compared with control patients. Among the 765 DEG identified in tubal epithelium, 512 genes were up-regulated and 253 genes were down-regulated in the TEM group. Validation using qPCR (P = 0.0288), immunohistochemistry and Western blotting (P = 0.0150) confirmed significant up-regulation of CCL2 expression in the TEM group. In-vitro experiments revealed that recombinant CCL2 protein significantly reduced CBF (100 ng/ml versus blank control: P < 0.0001), differentiation of ciliated cells (25 ng/ml versus blank control: P = 0.0206, 50 ng/ml versus blank control: P < 0.0001, 100 ng/ml versus blank control: P < 0.001) and ciliary length (P < 0.0001) in FTEC, suggesting a role for CCL2 in the development of the TEM-related pathological phenotype.
These findings indicate that CCL2 may contribute to the pathological phenotype associated with TEM, and could be a crucial signalling molecule in damage of tubal epithelium in patients with TEM.
输卵管子宫内膜异位症（TEM）如何影响输卵管上皮的形态和超微结构，以及TEM的潜在病理生理机制是什么？
从患有和未患有TEM的患者中收集输卵管上皮样本（TEM组，n = 27；对照组，n = 28）。通过苏木精和伊红染色、电子显微镜和高速摄像机评估形态学特征、超微结构、纤毛细胞百分比和纤毛摆动频率（CBF）。进行mRNA微阵列分析以鉴定输卵管上皮中差异表达的基因（DEG），并使用定量聚合酶链反应（qPCR）、免疫组织化学和蛋白质印迹进一步验证。在体外检测重组趋化因子配体2（CCL2）蛋白对原代人输卵管上皮细胞（FTEC）的影响。
与对照患者相比，TEM患者的输卵管上皮在形态和超微结构上表现异常，纤毛细胞百分比（P < 0.0001）和CBF（P < 0.0001）显著降低。在输卵管上皮中鉴定出的765个DEG中，TEM组中有512个基因上调，253个基因下调。使用qPCR（P = 0.0288）、免疫组织化学和蛋白质印迹（P = 0.0150）进行的验证证实TEM组中CCL2表达显著上调。体外实验表明，重组CCL2蛋白显著降低了FTEC中的CBF（100 ng/ml与空白对照相比：P < 0.0001）、纤毛细胞分化（25 ng/ml与空白对照相比：P = 0.0206，50 ng/ml与空白对照相比：P < 0.0001，100 ng/ml与空白对照相比：P < 0.001）和纤毛长度（P < 0.0001），表明CCL2在TEM相关病理表型的发展中起作用。
这些发现表明CCL2可能促成与TEM相关的病理表型，并且可能是TEM患者输卵管上皮损伤中的关键信号分子。
