Abstract
Leydig cell (LCs) apoptosis is responsible for decreased serum testosterone levels during late-onset hypogonadism (LOH). Our study was designed to illustrate the regulatory effect of lncRNA XIST on LCs and to clarify its molecular mechanism of action in LOH. The Leydig cells (TM3) was treated by 300 μM H2O2 for 8 h to establish Leydig cell oxidative stress model in vitro. The expression levels of lncRNA XIST in the testicular tissues of patients with LOH were measured using fluorescence in situ hybridization (FISH). The interaction between lncRNA XIST/SIRT1 and miR-145a-5p was assessed using starBase and dual-luciferase reporter gene assays. Apoptotic cells and Caspase3 activity were determined by flow cytometry (FCM) assay. Testosterone concentration was determined by ELISA. Moreover, histological assessment of testicles in mice was performed by using HE staining and the TUNEL assay was used to determine apoptosis. We found that the lncRNA XIST was downregulated in the testicular tissues of LOH patients and mice and in H2O2-induced TM3 cells. XIST siRNA significantly promoted apoptosis, enhanced Caspase3 activity and reduced testosterone levels in H2O2-stimulated TM3 cells. Further studies showed that the miR-145a-5p inhibitor reversed the effect of XIST-siRNA on H2O2-induced Leydig cell apoptosis. MiR-145a-5p negatively regulated SIRT1 expression, and SIRT1-siRNA reversed the effects of the miR-145a-5p inhibitor on H2O2 stimulated TM3 cells. The in vivo experiments indicated that silencing of the lncRNA XIST aggravated LOH symptoms in mice. Inhibition of lncRNA XIST induces Leydig cell apoptosis through the miR-145a-5p/SIRT1 axis in the progression of LOH.
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The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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This study was supported by The Application Value of Penile Neuroelectrophysiological Techniques in the Diagnosis and Treatment of Premature Ejaculation (grant no. BE2019606).
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Jing Wang, Yiqiong Yang, and Yang Xu contributed to data collection, statistical analysis, data interpretation and manuscript preparation. Zhipeng Xu and Xiaozhi Zhao contributed to data collection, statistical analysis. Ruipeng Jia and Yutian Dai contributed to data collection and manuscript preparation. All authors have read and approved the final manuscript.
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This study was approved by the Ethical committee of Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University. All animal experiments were conducted in accordance with the Animal Care and Use Committee of the Affiliated Nanjing Hospital of Nanjing Medical University.
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Wang, J., Yang, Y., Xu, Y. et al. Long noncoding RNA XIST inhibition promotes Leydig cell apoptosis by acting as a competing endogenous RNA for microRNA-145a-5p that targets SIRT1 in late-onset hypogonadism. Cell Tissue Res 395, 285–297 (2024). https://doi.org/10.1007/s00441-024-03860-3
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DOI: https://doi.org/10.1007/s00441-024-03860-3