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Abstract
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Non-obstructive azoospermia (NOA) is a severe form of male infertility, yet its underlying molecular mechanisms
remain poorly understood. This study aimed to identify key regulatory non-coding RNAs (ncRNAs) and hub
genes associated with NOA through an integrative bioinformatics approach. Using microarray analysis, we
examined 4956 ncRNAs and identified 29 differentially expressed ncRNAs (14 upregulated, 15 downregulated)
in NOA compared to healthy individuals. Co-expression analysis revealed significant interactions between
lncRNAs, miRNAs, and mRNAs, predicting 31 target mRNAs within the regulatory network. Further, single-cell
transcriptomic analysis identified four pivotal hub genes in NOA Sertoli cells: CLTC, XIAP, and DHFR (upregulated)
and STMN1 (downregulated). Functional enrichment analysis highlighted critical pathways, including
mitotic spindle organization and phosphatase activity, suggesting their involvement in NOA pathophysiology.
Our findings provide novel insights into the molecular mechanisms underlying NOA and propose potential
biomarkers for improved diagnosis and therapeutic strategies.
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