Seminal Plasma Exosomal miRNA Profiling Reveals hsa‐miR‐7‐5p as a Key Regulator of Sperm Motility in Asthenozoospermia

BACKGROUND: Asthenozoospermia is a significant contributing factor to male infertility. Accumulating evidence indicates that impaired sperm motility is closely linked to dysregulated microRNA expression during spermatogenesis. Seminal plasma exosomes are enriched with diverse microRNAs, which play pivotal roles in modulating sperm motility, the acrosome reaction, capacitation, and fertilization. OBJECTIVE: To investigate the association between exosomal microRNA profiles and sperm motility by comparing microRNA expression patterns in seminal plasma exosomes from asthenozoospermic patients and healthy controls, thereby elucidating the molecular mechanisms underlying exosomal microRNA-mediated regulation of sperm motility. MATERIALS AND METHODS: Seminal plasma exosomes were isolated from 21 asthenozoospermic patients and 21 age-matched healthy donors via ultracentrifugation, followed by characterization via transmission electron microscopy, nanoparticle tracking analysis, and Western blotting. Exosomal microRNA profiles were analyzed via unique molecular identifier-based small RNA sequencing to identify motility-associated microRNAs. RESULTS: Comparative analysis revealed 13 significantly upregulated microRNAs in the exosomes of asthenozoospermic patients, among which hsa-miR-7-5p exhibited the greatest abundance. Functional enrichment analysis revealed that the target genes of hsa-miR-7-5p were associated primarily with cellular metabolism, macromolecule metabolic processes, and nitrogen compound metabolism, as well as several crucial signaling pathways, including mTOR signaling, endocrine resistance, and gonadotropin-releasing hormone secretion. A dual-luciferase reporter assay verified that hsa-miR-7-5p directly targets rapidly accelerated fibrosarcoma 1, a pivotal component of the gonadotropin-releasing hormone signaling pathway. DISCUSSION AND CONCLUSION: hsa-miR-7-5p may regulate sperm motility through the rapidly accelerated fibrosarcoma 1 -mediated gonadotropin-releasing hormone secretion and MAPK signaling networks. Our findings highlight the potential role of exosomal microRNAs in male infertility. Future prospective studies are warranted to elucidate the molecular mechanisms underlying the regulation of sperm motility.