International Journal of Molecular Sciences · Nov 3, 2026
Single-Cell Transcriptomic Analysis of Rat Urethral Tissue Following Simulated Childbirth Injury and Microenergy Acoustic Pulse Therapy
By Lin G, Zhao L, Xing E et al.
Microenergy acoustic pulse (MAP) therapy has been associated with functional and structural improvement after childbirth-related urethral injury, but its multicellular effects remain incompletely defined. We explored treatment- and injury-associated differences in the urethral microenvironment using single-cell RNA sequencing. Eight female Sprague–Dawley rats were assigned to sham, vaginal balloon dilation plus ovariectomy (VBDO), VBDO plus beta-aminopropionitrile (BAPN), or VBDO plus BAPN and MAP (MAP) groups, with one 24-week-old and one 48-week-old rat per group. Urethral tissues were analyzed for cell composition, differential gene expression, Gene Ontology enrichment, stem/progenitor features, senescence-associated secretory phenotype activity, and CellChat-inferred intercellular communication. A balanced single-cell dataset comprising 23,128 cells was used for downstream comparative analyses and resolved 19 annotated cell-type/state categories. Across the injury- and age-associated experimental conditions, epithelial, vascular, stromal, immune, and muscle compartments showed distinct transcriptional patterns, whereas MAP-treated samples showed a distinct cellular distribution; a separate histological cohort showed higher urethral muscle content. MAP-treated samples also showed differences in CellChat-inferred signaling activity in selected extracellular-matrix, adhesion, vascular, developmental, and neural/neuromuscular pathways, including fibronectin 1 (FN1), collagen (COLLAGEN), angiopoietin (ANGPT), vascular endothelial growth factor (VEGF), Wingless/Integrated (WNT), Notch (NOTCH), reelin (RELN), Slit guidance ligand (SLIT), neural cell adhesion molecule (NCAM), and neurexin (NRXN). Exploratory enrichment analysis identified treatment-associated differences in translation, oxidative metabolism, stress adaptation, synaptic-organization processes, progenitor-associated transcriptional states, and senescence-related programs. The treatment-split marker manifold approximation and projection (UMAPs) are descriptive and do not provide evidence of increased muscle-cell number; the reported difference in muscle-associated staining content derives from a separate animal-level histological analysis. Overall, the findings are descriptive and hypothesis-generating and require validation in independently replicated cohorts.