Zhenwu Decoction ameliorates experimental benign prostatic hyperplasia: association with mitochondrial quality...

Zhenwu Decoction (ZWD), first recorded in the Shang Han Lun, is a classical formula used to warm Yang and promote water metabolism. It has traditionally been used for Yang deficiency with water retention, impaired Qi transformation and dysuria. Benign prostatic hyperplasia (BPH) can be classified under the traditional categories of "Longbi" and "dysuria", and its pathogenesis is closely related to kidney Yang deficiency, impaired bladder Qi transformation and water retention in the lower Jiao. Given that BPH is not merely a proliferative lesion but a chronic process involving androgenic stimulation, oxidative stress, insufficient apoptosis and tissue remodelling, investigating the modern pharmacological basis by which ZWD improves BPH through multiple pathological links has clear ethnopharmacological relevance.

This study aimed to evaluate the therapeutic effect of ZWD on TP-induced experimental BPH and to determine whether this effect is accompanied by changes in oxidative stress, apoptosis and markers of HIF-1α/BNIP3-related mitochondrial quality control.

UPLC-MS/MS was used to characterise the chemical constituents of ZWD, and network pharmacology analysis was performed based on the experimentally detected candidate compounds. A testosterone propionate (TP)-induced rat model of BPH was established to evaluate the effects of ZWD on prostatic hyperplasia, tissue remodelling, androgen levels, oxidative stress and apoptosis. Western blotting, qPCR, immunohistochemistry, immunofluorescence co-localisation, mCherry-GFP-LC3 autophagy flux-related assays and medicated serum experiments in BPH-1 cells were further used to assess changes associated with mitochondrial quality control.

UPLC-MS/MS combined with database cross-matching identified 49 ZWD-related candidate compounds, of which 24 met the criteria of OB ≥ 20% and DL ≥ 0.10. Network pharmacology analysis identified 135 overlapping ZWD-BPH targets. These targets were mainly enriched in processes related to the MAPK cascade, reactive oxygen species response, AGE-RAGE signalling, HIF-1 signalling, EGFR-related signalling and endocrine regulation. In vivo, ZWD reduced prostate volume, prostate weight and prostate index in TP-induced BPH rats. It also improved epithelial thickening, glandular architectural disruption and collagen deposition, decreased serum T, DHT and prostatic MDA levels, and increased SOD activity. ZWD treatment also enhanced TUNEL-positive signals, up-regulated Bax and cleaved caspase-3, down-regulated Bcl-2, and was accompanied by changes in mitochondrial quality control-related markers, including HIF-1α, BNIP3, Beclin-1, LC3B-II/LC3B-I and p62. In vitro, ZWD-medicated serum reduced BPH-1 cell viability and ROS accumulation, improved mitochondrial membrane potential status, promoted apoptosis-related changes, and enhanced the spatial association between mitochondria and the autophagy-lysosome system.

ZWD ameliorated prostatic hyperplasia and tissue remodelling in experimental BPH. These effects were accompanied by reduced androgenic stimulation, decreased oxidative stress, enhanced apoptosis-related changes and restoration of mitochondrial quality control-related markers. HIF-1α/BNIP3-related mitochondrial quality control may represent an important observational node in the action of ZWD, although its causal role requires further clarification through targeted intervention and dynamic flux validation.