Qiu Yingying, Wei Xinjun, Cao Jian, Wang Jindan, Dou Qianru, Zhou Fangfang, Chen Xi, Liu Yongli, Wan Guiping, Huang Meihua, Li Zhenli, Gui Tao
Department of Obstetrics and Gynecology, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, China.
Department of Gynecology, The Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou, 221112, China.
Reprod Sci. 2025 Aug 6. doi: 10.1007/s43032-025-01944-1.
Adenomyosis (AM), a gynecological disorder that severely affects female reproductive health. AM-associated macrophage (AAM) polarization-induced epithelial-mesenchymal transition (EMT) is a key driver of AM progression. In this study, we investigated the role and underlying mechanisms of endometrial mesenchymal stem cell (eMSC)-derived exosomes in regulating AAM polarization and the subsequent EMT of endometrial epithelial cells (EECs). In vitro coculture studies revealed that AM eutopic eMSCs markedly induced M2 macrophage polarization via exosomes and promoted EMT of EECs. Differentially expressed microRNAs (DE-miRNAs) between exosomes derived from normal eMSCs (N-eMSCs) and AM eutopic eMSCs (A-eMSCs) were identified using miRNA sequencing and miR-4669 was found to be the most significantly upregulated miRNA. Internalization of exosomal miR-4669 by macrophages induced their polarization toward the M2 phenotype and promoted the EMT of EECs. Mechanistic analysis using luciferase assay, mRNA sequencing, and rescue experiments revealed that miR-4669 induced M2 macrophage polarization via downregulation of DUSP6 and activation of MAPK/ERK signaling. The polarized M2 macrophages promoted the EMT of ISK cells via TGF-β1 secretion. In an AM xenograft mouse model, miR-4669 depletion inhibited AM progression by targeting the DUSP6/ERK1/2 pathway in macrophages. Overall, AM A-eMSC-derived exosomal miR-4669 facilitates M2 macrophage polarization by targeting the DUSP6/ERK signaling pathway, thereby promoting EMT of EECs via TGF-β1 secretion. These findings open avenues for developing novel preventive and therapeutic strategies for AM.
子宫腺肌病（AM）是一种严重影响女性生殖健康的妇科疾病。AM相关巨噬细胞（AAM）极化诱导的上皮-间质转化（EMT）是AM进展的关键驱动因素。在本研究中，我们探讨了子宫内膜间充质干细胞（eMSC）衍生的外泌体在调节AAM极化及随后子宫内膜上皮细胞（EEC）的EMT中的作用和潜在机制。体外共培养研究表明，AM在位eMSC通过外泌体显著诱导M2巨噬细胞极化，并促进EEC的EMT。使用miRNA测序鉴定了正常eMSC（N-eMSC）和AM在位eMSC（A-eMSC）衍生的外泌体之间差异表达的微小RNA（DE-miRNA），发现miR-4669是上调最显著的miRNA。巨噬细胞对内源性miR-4669的内化诱导其向M2表型极化，并促进EEC的EMT。使用荧光素酶测定、mRNA测序和拯救实验进行的机制分析表明，miR-4669通过下调DUSP6和激活MAPK/ERK信号通路诱导M2巨噬细胞极化。极化的M2巨噬细胞通过分泌TGF-β1促进ISK细胞的EMT。在AM异种移植小鼠模型中，miR-4669缺失通过靶向巨噬细胞中的DUSP6/ERK1/2途径抑制AM进展。总体而言，AM A-eMSC衍生的外泌体miR-4669通过靶向DUSP6/ERK信号通路促进M2巨噬细胞极化，从而通过分泌TGF-β1促进EEC的EMT。这些发现为开发AM的新型预防和治疗策略开辟了道路。
