Molecular mechanisms and diagnostic value of seminal plasma exosomal miR-26a-5p targeting PTEN in idiopathic teratozoospermia
LIU Chunhui
WU Ruipeng
WANG Zhiqiang
SHAN Wensheng
LI Shaojun
Abstract：Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.
Keywords：seminal plasma exosomesteratozoospermiamiR-26a-5pPTENmolecular mechanisms
Publication Date：2025-10-25
Online Publishing Date：2025-11-05(First online date of this platform, not the publication date of the document)
Pages：11( 3256-3266 )