Li Li, Wang Xiaoyan, Wu You, Liu Yi, Zhang Mingzhe
Departnent of Reproductive Medicine, Affiliated Hospital of Zunyi Medical University, No. 149 Dalian Road, Zunyi, Guizhou Province, 563000, China.
Departnent of Gyneccology, Affiliated Hospital of Zunyi Medical University, No. 149 Dalian Road, Zunyi, Guizhou Province, 563000, China.
Appl Biochem Biotechnol. 2025 Jun 23. doi: 10.1007/s12010-025-05290-5.
Endometriosis (EMs) is a chronic inflammatory disorder featured by infertility and pain. The role of N6-methyladenosine (m6A) in EMs has been evidenced. This study investigated the molecular mechanism of METTL14-m6A-KLF4 modulating macrophage polarization in EMs. RT-qPCR assay was conducted to test METTL14 levels in tissues and cells and the relative mRNA levels of M2 (Arg-1, Fizz1) and M1 (iNOS, IL-1β) factors in the supernatant after co-culture of macrophages with normal endometrial stromal cells (nESCs) or ectopic endometrial stromal cells (eESCs). CD206 and CD86 expression, as well as Arg-1, IL-10, and IL-4 levels, were assessed. Meanwhile, the relationship between METTL14 and the m6A modification of KLF4 was analyzed. Additionally, the effect of KLF4-activated M2 macrophages on in vitro ESC progression was observed. Cellular and tissue METTL14 was under-expressed in EMs. METTL14 expression might be related to macrophage M2 polarization. Co-culture of eESCs overexpressing METTL14 and macrophages downregulated Arg-1, Fizz1, CD206, IL-10, and IL-4 levels. Mechanistically, METTL14 could mediate KLF4 m6A modification through the m6A reading protein YTHDF2. KLF4 overexpression could nullify METTL14 re-expression-repressed M2 macrophage polarization. In addition, KLF4-activated M2 macrophages accelerated the proliferation and migration of ESCs in vitro. METTL14-m6A-KLF4 regulated macrophage polarization in EMs.
子宫内膜异位症（EMs）是一种以不孕和疼痛为特征的慢性炎症性疾病。N6-甲基腺苷（m6A）在EMs中的作用已得到证实。本研究探讨了METTL14-m6A-KLF4调节EMs中巨噬细胞极化的分子机制。采用RT-qPCR检测组织和细胞中METTL14水平，以及巨噬细胞与正常子宫内膜基质细胞（nESCs）或异位子宫内膜基质细胞（eESCs）共培养后上清液中M2（Arg-1、Fizz1）和M1（iNOS、IL-1β）因子的相对mRNA水平。评估CD206和CD86表达以及Arg-1、IL-10和IL-4水平。同时，分析METTL14与KLF4的m6A修饰之间的关系。此外，观察KLF4激活的M2巨噬细胞对体外ESC进展的影响。细胞和组织中的METTL14在EMs中表达下调。METTL14表达可能与巨噬细胞M2极化有关。过表达METTL14的eESCs与巨噬细胞共培养可下调Arg-1、Fizz1、CD206、IL-10和IL-4水平。机制上，METTL14可通过m6A阅读蛋白YTHDF2介导KLF4的m6A修饰。KLF4过表达可抵消METTL14重新表达抑制的M2巨噬细胞极化。此外，KLF4激活的M2巨噬细胞在体外加速了ESCs的增殖和迁移。METTL14-m6A-KLF4调节EMs中的巨噬细胞极化。
