Decoding the ceRNA regulatory landscape: a comprehensive transcriptomic analysis of ncRNA-driven bladder fibrosis following spinal cord injury

解码ceRNA调控图谱:脊髓损伤后ncRNA驱动的膀胱纤维化的全面转录组分析

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Abstract

BACKGROUND: Neurogenic bladder (NB) following spinal cord injury (SCI) is a debilitating complication characterized by bladder fibrosis and structural remodeling. Non-coding RNAs (ncRNAs) are emerging regulators of fibrotic processes, yet their roles in SCI-induced NB remain unexplored. This study aimed to delineate the dynamic ncRNA-messenger RNA (mRNA) regulatory landscape in SCI-associated NB using transcriptomic profiling, identifying potential biomarkers and therapeutic targets. METHODS: Twenty female Wistar rats underwent complete T10-T11 spinal cord transection (SCI groups, n=15) or laminectomy-only (NC group, n=5). Bladder tissues were harvested at 1, 2, and 4 weeks post-SCI. Next-generation sequencing (NGS) analyzed mRNA, long non-coding RNA (lncRNA), and microRNA (miRNA) expression. Differentially expressed genes/mRNAs/lncRNAs/miRNAs (DEGs/DEMs/DELs/DEMIs) were identified (|log2FC|>1, P<0.05), and lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks were constructed. Functional enrichment and quantitative real-time polymerase chain reaction (qPCR) validation were performed to confirm key findings. RESULTS: Histopathology confirmed progressive bladder fibrosis, with collagen deposition increasing from 7.3% (NC) to 34.6% at 4 weeks post-SCI (P<0.001). NGS revealed 3,255/3,449/884 DEMs, 904/870/278 DELs, and 229/77/127 DEMIs in SCI-1/2/3 vs. NC, respectively. Temporal analysis identified 420 shared DEMs and 102 DELs, with lncRNAs enriched on chromosomes 1 and 7. The ceRNA networks implicated transforming growth factor-beta (TGF-β), calcium, and interleukin-17 (IL-17) signaling pathways in fibrosis. Notably, miR-21-5p showed progressive upregulation, while miR-139-5p decreased, correlating with ROCK2-mediated detrusor overactivity (DO). Functional enrichment highlighted inflammatory response and extracellular matrix reorganization as dominant processes. qPCR validated 12 candidate DEGs, including PVT1 and miR-21-5p, aligning with sequencing data. CONCLUSIONS: This first comprehensive transcriptomic atlas of SCI-induced NB uncovers dynamic ncRNA-mRNA networks driving fibrosis through TGF-β and inflammatory pathways. The identified dysregulated ncRNAs and their target genes provide novel biomarkers for early NB diagnosis and potential therapeutic entry points. Our findings establish a foundation for developing ncRNA-targeted strategies to mitigate bladder remodeling post-SCI, addressing a critical unmet need in neuro-urology.

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