Exploring the potential mechanism of Huang'e capsule against spontaneous benign prostatic hyperplasia in beagle dogs using high-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry, gas chromatography-mass spectrometry, and network pharmacology.

[OBJECTIVE] To investigate the therapeutic efficacy and potential mechanisms of Huang'e capsule (, HEC) against benign prostatic hyperplasia (BPH).

[METHODS] The chemical profile of HEC was characterized using high-performance liquid chromatographyquadrupole-time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) and gas chromatography-mass spectrometry (GC-MS) techniques. Network pharmacology was employed to analyze potential active compounds, core targets, and key signaling pathways. A spontaneous canine BPH model was used to evaluate the efficacy of HEC and to validate the predictions from network pharmacology.

[RESULTS] A total of 51 chemical components of HEC were identified, comprising 19 from HPLC-Q-TOF-MS/MS and 32 from GC-MS analyses. The "components-targets-pathways-disease" network analysis predicted active compounds including (s)-coriolic acid, ethyl linoleate, peroxysimulenoline, physcion, and kaempferol. Core targets identified included cytochrome P450 family 19 subfamily A member 1, estrogen receptor 2 (ESR2), ESR1, and androgen receptor (AR). Kyoto Encyclopedia of Genes and Genomes enrichment analysis suggested that HEC's effects on BPH involve pathways related to cancer, phosphatidylinositol 3-kinase (PI3K) -protein kinase B (Akt)-signaling, proteoglycans in cancer, and prostate cancer signaling. Animal experiments showed that HEC significantly improved maximum urinary flow rates, reduced prostate weight, volume, and prostate index, and ameliorated histopathological changes. HEC regulated the balance between apoptosis and proliferation by downregulating AR and estrogen receptor alpha expression, while upregulating estrogen receptor beta expression.

[CONCLUSION] These findings indicate that HEC effectively ameliorates spontaneous BPH in beagle dogs, likely by regulating the balance between cell apoptosis and proliferation through multiple signaling pathways.