Bladder outlet obstruction (BOO), commonly resulting from conditions such as benign prostatic hyperplasia, leads to significant bladder dysfunction, including detrusor underactivity (DU). Although mesenchymal stem cell (MSC) therapy has demonstrated therapeutic potential, the underlying molecular mechanisms remain unclear. Human umbilical cord-derived MSCs (hUCMSCs) offer advantages such as low immunogenicity, easy accessibility, and potent regenerative capabilities.
This study aims to integrate proteomic and transcriptomic datasets to elucidate the molecular mechanisms of hUCMSC therapy. We hypothesized that combining both omics approaches would provide a comprehensive view of the pathways and molecular targets involved in bladder repair following BOO.

Rats were assigned to sham, BOO, and BOO + hUCMSC treatment groups. The hUCMSC-treated group received 2×10⁶ cell doses. Urodynamic studies and histological analyses were conducted on day 56. For proteomics, bladder tissues were analyzed via LC-MS/MS, followed by GO and KEGG enrichment analyses. RNA-seq was performed on bladders from sham, BOO, and BOO + hUCMSC rats, with data analyzed using clusterProfiler and STRINGdb for pathway and PPI analysis.

Urodynamically and histologically, the study confirmed that hUCMSC treatment significantly improved bladder function and reversed histological damage in a dose-dependent manner.
BOO altered expression of 1,536 mRNAs (987 upregulated, 549 downregulated), and hUCMSC treatment reversed many of these changes. GO analysis indicated involvement in inflammatory response and oxygen transport. KEGG analysis highlighted cytokine-cytokine receptor interaction and IL-17 signaling pathways. Spp1 was a key gene downregulated after hUCMSC treatment.
Proteomic profiling identified 2,510 proteins, with 2,452 statistically analyzed. KEGG enrichment in the hUCMSC vs. BOO group revealed key pathways including PI3K-Akt, AMPK, Hippo signaling, and cell cycle regulation—corroborating transcriptomic results.

The study supports the anti-inflammatory and anti-fibrotic effects of hUCMSCs. Notably, transcriptomic data highlight Spp1 as a candidate molecular target, involved in tissue remodeling and inflammation, with expression reduced by hUCMSC therapy. Proteomic data emphasize the role of PI3K-Akt and AMPK signaling, both critical in cell survival and metabolism.
The convergence of these findings reinforces the therapeutic potential of hUCMSCs and provides a dual-level mechanistic framework for understanding bladder regeneration. Importantly, Spp1 and PI3K-Akt may act in concert or independently to mediate therapeutic benefits. However, further investigations between SPP1 and the PI3K-Akt and AMPK signaling pathway in BOO-Induced bladder dysfunction are needed.

This study reveals that hUCMSCs mitigate BOO-induced bladder dysfunction via regulation of inflammatory pathways, extracellular matrix remodeling, and critical signaling cascades such as PI3K-Akt and AMPK. Spp1 emerges as a novel target gene, providing further insight into the therapeutic mechanism and potential for future drug development.