Fang Xingyu, Lu Xiaotong, Ma Yujie, Sun Ning, Jiao Yunyun, Meng Hui, Song Mengjiao, Jin Haixia, Yao Guidong, Song Ning, Wu Zhaoting, Wen Shuang, Guo Haoran, Xiong Haosen, Song Wenyan
Center for Reproductive Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Department of Pathology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
iScience. 2024 Sep 10;27(10):110904. doi: 10.1016/j.isci.2024.110904. eCollection 2024 Oct 18.
Non-obstructive azoospermia (NOA) is the most incurable form of male infertility with a complex etiology. Long non-cording RNAs (lncRNAs) were associated with regulating spermatogenesis. Herein, differentially expressed lncRNAs between NOA and control male were screened by RNA-seq analysis. MEG3 was upregulated in NOA tissues and inhibited cell proliferation and promoted cell autophagy and apoptosis . Through RNA immunoprecipitation (RIP), biotin pull-down assays, and dual-luciferase reporter assays, MEG3 was proved to act as a competing endogenous RNA of microRNA (miR)-21 and thus influenced the SPRY1/ERK/mTOR signaling pathway. Additionally, bioinformatic prediction and chip assay revealed that MEG3 was possibly regulated by nuclear factor κB (NF-κB) and SPRY1/NF-κB/MEG3 formed a feedback loop. Seminiferous tubule microinjection further investigated the effects of MEG3 on testes . These findings demonstrated that MEG3-miR-21-SPRY1-NF-κB probably acted as a feedback loop leading to azoospermia. Our study might provide a target and theoretical basis for diagnosing and treating NOA.
非梗阻性无精子症（NOA）是男性不育中最难治愈的类型，其病因复杂。长链非编码RNA（lncRNAs）与精子发生的调控有关。在此，通过RNA测序分析筛选出NOA男性与对照男性之间差异表达的lncRNAs。MEG3在NOA组织中上调，抑制细胞增殖，促进细胞自噬和凋亡。通过RNA免疫沉淀（RIP）、生物素下拉试验和双荧光素酶报告试验，证明MEG3作为微小RNA（miR）-21的竞争性内源性RNA，从而影响SPRY1/ERK/mTOR信号通路。此外，生物信息学预测和芯片分析表明，MEG3可能受核因子κB（NF-κB）调控，且SPRY1/NF-κB/MEG3形成一个反馈环。曲细精管显微注射进一步研究了MEG3对睾丸的影响。这些发现表明，MEG3-miR-21-SPRY1-NF-κB可能作为导致无精子症的反馈环。我们的研究可能为NOA的诊断和治疗提供靶点和理论依据。
