Kangquan formula ameliorates the benign prostatic hyperplasia induced by testosterone propionate in rats by regulating urine microecology and metabolic profile.

Background: Kangquan formula (KQF), a traditional Chinese medicine compound, is clinically effective and safe against benign prostatic hyperplasia (BPH). However, the mechanism of action of KQF is not clear.

Objective: To explore the potential mechanism of action of KQF on BPH by integrating urine microecology and metabolomics.

Methods: The BPH rat model was induced using testosterone propionate. Histopathology, immunofluorescence, and immunohistochemistry staining were used to evaluate the curative effect of KQF on BPH. In addition, 16S rRNA sequencing, nontargeted metabolomics, and western blot analysis were performed to analyze the effects of KQF on the urinary microecological diversity, metabolic profile, and epithelial barrier in rats with BPH. Finally, association analysis was used to demonstrate the correlation among urinary microorganisms, metabolites, and phenotypes.

Results: KQF decreased the wet weight, volume, and prostate index in rats with BPH, promoted the expression of caspase-3, a marker of apoptosis in prostate tissue, and inhibited the expression of proliferation cell nuclear antigen (PCNA), a marker of proliferation. Low-dose KQF intervention increased the urine flora richness, reduced the abundance of harmful bacteria such as Morganella, and increased the abundance of probiotics such as Staphylococcus, Romboutsia, Turicibacter, and Bacillus. In addition, low-dose KQF intervention increased the content of 103 metabolites, including citramalic acid and uracil, while also upregulating the content of 120 metabolites, including pralidoxime and d-glucuronic acid, in the urine of rats with BPH. Glycerophospholipid, glycine, serine, threonine, nucleotide, amino sugar, and nucleotide sugar may be the key metabolic pathways involved in the therapeutic effects of KQF. Moreover, low-dose KQF intervention upregulated the protein expression of uroepithelial barrier markers ZO-1, occludin, and UP II in the prostatic area of rats with BPH. The association analysis revealed overall consistency and differential correlation among urine microbes, metabolites, and phenotypes.

Conclusions: This study was the first to analyze the mechanism of action of KQF on BPH from the perspective of urine microenvironment. KQF alleviated the pathological changes of prostate tissue in rats with BPH and regulated the imbalance of cell proliferation/apoptosis in BPH tissues. It also exerted its effect probably by improving the diversity of urine microecology, restoring urinary metabolic profile homeostasis, and remodeling the integrity of the urinary epithelial barrier in rats with BPH.