Didang Tang downregulates prostatic necroptosis and NETs formation in rats with chronic prostatitis: Evidence from multi-omics analysis and experimental validation.

ETHNOPHARMACOLOGICAL RELEVANCE: Didang Tang (DDT), a classic traditional Chinese medicine formula, possesses the effects of activating blood circulation, resolving stasis, reducing swelling, and alleviating pain. It has demonstrated significant therapeutic efficacy in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS), effectively relieving clinical symptoms and improving remission rates. However, its underlying molecular mechanisms remain incompletely elucidated.

AIM OF THE STUDY: The present study aimed to investigate the therapeutic effects of DDT on CP/CPPS and to elucidate the potential mechanisms by which it ameliorates prostatic tissue injury.

MATERIALS AND

METHODS

A CFA-induced CP/CPPS-like prostatitis rat model was established by intra-prostatic injection of Freund's complete adjuvant in male Sprague-Dawley (SD) rats. DDT was administered by oral gavage to evaluate its therapeutic efficacy. Following tissue collection, proteomic and transcriptomic analyses were performed to explore the molecular mechanisms of DDT action. Single-cell RNA sequencing (scRNA-seq) was employed to investigate the causal relationship between necroptosis and CP/CPPS, thereby laying the foundation for clarifying the complex pathogenesis of this disease. Histological changes in prostatic tissue, levels of necroptosis, NETs formation, and expression of the RIPK3/MLKL signaling pathway were assessed using hematoxylin-eosin (HE) staining, immunohistochemistry (IHC), immunofluorescence (IF), Western blot.

RESULTS

DDT effectively reduced the level of necroptosis, decreased NETs formation, and improved histopathological injury in the CFA-induced CP/CPPS-like rat model. Proteomic and transcriptomic analyses further suggested that the protective effects of DDT in this prostatitis-like inflammatory injury model may be associated with the regulation of necroptosis-related pathways and NETs formation.

CONCLUSIONS

This study provides initial experimental evidence showing that representative necroptosis-related pathway markers and NETs-associated markers are upregulated in a CFA-induced CP/CPPS-like animal model. DDT treatment dose-dependently attenuated the expression of these markers and improved prostatitis-like histopathological injury. These findings suggest that suppression of necroptosis-associated HMGB1 release and NETs formation may represent one possible mechanism by which DDT exerts its protective effects in prostatitis-like inflammatory injury. This study provides supportive evidence for the potential application of traditional Chinese medicine in prostatitis-like inflammatory conditions and offers a basis for further mechanistic and therapeutic studies.