Abstract：
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.
摘要：
子宫腺肌病的发病机制与上皮间质转化和巨噬细胞密切相关。已经广泛研究了MicroRNA与一系列恶性肿瘤中上皮-间质转化的关系。然而,关于子宫腺肌病在位子宫内膜的细胞外囊泡及其包裹的microRNAs的研究很少。在这项研究中,我们研究了源自细胞外囊泡的microRNA-25-3p在子宫腺肌病患者和对照组子宫内膜上皮细胞中诱导巨噬细胞极化和促进上皮-间质转化中的作用.我们从原代子宫内膜细胞培养上清液中获得了在位子宫内膜样品和分离的细胞外囊泡。实时定量PCR分析表明microRNA-25-3p在细胞外囊泡中高表达,以及在由子宫腺肌病在位子宫内膜的细胞外囊泡刺激的巨噬细胞中；用microRNA-25-3p转染的巨噬细胞表现出升高的M2标记水平，同时显示降低的M1标记水平。与上述极化巨噬细胞共培养后，子宫内膜上皮细胞表达较高水平的N-cadherin和波形蛋白，E-cadherin和细胞角蛋白7的蛋白质水平较低。结果表明，包裹在在位子宫内膜细胞胞外囊泡中的microRNA-25-3p可以诱导巨噬细胞向M2极化，极化的巨噬细胞促进上皮细胞的上皮-间质转化。然而,体外实验显示子宫腺肌病组与对照组子宫内膜上皮细胞的迁移能力无明显差异。此外，观察到microRNA-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-间质转化和迁移。因此，microRNA-25-3p诱导的极化巨噬细胞在促进子宫腺肌病发展中的意义尚不清楚，和巨噬细胞浸润单独可能是足够的这个过程。我们强调局部在位子宫内膜微环境的特异性，并假设其在子宫腺肌病发病机理中的潜在意义。
