Department of Gynecology and Obstetrics, Qilu Hospital (Qingdao), Cheeloo College of Medicine, Shandong University, Qingdao, China.
Department of Obstetrics and Gynecology, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, China.
Mol Hum Reprod. 2024 Mar 28;30(4). doi: 10.1093/molehr/gaae010.
The pathogenesis of adenomyosis is closely related to the epithelial-mesenchymal transition and macrophages. MicroRNAs have been extensively investigated in relation to the epithelial-mesenchymal transition in a range of malignancies. However, there is a paucity of research on extracellular vesicles derived from the eutopic endometrium of adenomyosis and their encapsulated microRNAs. In this study, we investigated the role of microRNA-25-3p derived from extracellular vesicles in inducing macrophage polarization and promoting the epithelial-mesenchymal transition in endometrial epithelial cells of patients with adenomyosis and controls. We obtained eutopic endometrial samples and isolated extracellular vesicles from the culture supernatant of primary endometrial cells. Real-time quantitative PCR analysis demonstrated that microRNA-25-3p was highly expressed in extracellular vesicles, as well as in macrophages stimulated by extracellular vesicles from eutopic endometrium of adenomyosis; and macrophages transfected with microRNA-25-3p exhibited elevated levels of M2 markers, while displaying reduced levels of M1 markers. After co-culture with the above polarized macrophages, endometrial epithelial cells expressed higher levels of N-cadherin and Vimentin, and lower protein levels of E-cadherin and Cytokeratin 7. It was revealed that microRNA-25-3p encapsulated in extracellular vesicles from eutopic endometrial cells could induce macrophage polarization toward M2, and the polarized macrophages promote epithelial-mesenchymal transition in epithelial cells. However, in vitro experiments revealed no significant disparity in the migratory capacity of endometrial epithelial cells between the adenomyosis group and the control group. Furthermore, it was observed that microRNA-25-3p-stimulated polarized macrophages also facilitated the epithelial-mesenchymal transition and migration of endometrial epithelial cells within the control group. Thus, the significance of microRNA-25-3p-induced polarized macrophages in promoting the development of adenomyosis is unclear, and macrophage infiltration alone may be adequate for this process. We emphasize the specificity of the local eutopic endometrial microenvironment and postulate its potential significance in the pathogenesis of adenomyosis.
腺肌病的发病机制与上皮-间充质转化和巨噬细胞密切相关。微小 RNA 已广泛应用于多种恶性肿瘤的上皮-间充质转化研究。然而，关于源自腺肌病在位子宫内膜的细胞外囊泡及其包裹的微小 RNA 的研究却很少。在这项研究中，我们研究了源自细胞外囊泡的微小 RNA-25-3p 在诱导巨噬细胞极化和促进腺肌病患者和对照患者子宫内膜上皮细胞上皮-间充质转化中的作用。我们获得了在位子宫内膜样本，并从原代子宫内膜细胞的培养上清液中分离出细胞外囊泡。实时定量 PCR 分析表明，微小 RNA-25-3p 在细胞外囊泡中以及在由腺肌病在位子宫内膜来源的细胞外囊泡刺激的巨噬细胞中高度表达；转染微小 RNA-25-3p 的巨噬细胞表现出更高水平的 M2 标志物，而 M1 标志物水平降低。与上述极化的巨噬细胞共培养后，子宫内膜上皮细胞表达更高水平的 N-钙黏蛋白和波形蛋白，而 E-钙黏蛋白和细胞角蛋白 7 的蛋白水平降低。结果表明，源自在位子宫内膜细胞的细胞外囊泡中包裹的微小 RNA-25-3p 可诱导巨噬细胞向 M2 极化，极化的巨噬细胞促进上皮细胞的上皮-间充质转化。然而，在体外实验中，腺肌病组和对照组子宫内膜上皮细胞的迁移能力没有显著差异。此外，观察到微小 RNA-25-3p 刺激的极化巨噬细胞也促进了对照组子宫内膜上皮细胞的上皮-间充质转化和迁移。因此，微小 RNA-25-3p 诱导的极化巨噬细胞在促进腺肌病发展中的意义尚不清楚，巨噬细胞浸润本身可能足以完成这一过程。我们强调了局部在位子宫内膜微环境的特异性，并推测其在腺肌病发病机制中的潜在意义。
