Investigating the potential active components and mechanisms of Xiongcan Yishen formula in treating late-onset hypogonadism using network pharmacology combined with metabolomics and absorbed components.

ETHNOPHARMACOLOGICAL RELEVANCE: Xiongcan Yishen formula (XCYSF) is a traditional Chinese medicine (TCM) formula used for the treatment of male late-onset hypogonadism (LOH). However, its active components and underlying mechanisms remain unclear.

AIM OF THE STUDY: To investigate the potential pharmacodynamic material basis and mechanisms of XCYSF against LOH by integrating serum pharmacochemistry, network pharmacology, and metabolomics.

MATERIALS AND

METHODS

The blood-entry components of XCYSF in rat serum were identified using UPLC-Q-TOF-MS. Network pharmacology was employed to predict the targets of these components as well as LOH-related targets. Metabolomics was performed to analyze differential serum metabolites in a cyclophosphamide-induced LOH rat model. Integrated analyses were conducted to identify core targets and pathways, which were further validated by molecular docking. In vivo experiments were carried out using a LOH rat model, and in vitro experiments were performed using H2O2-induced TM3 cells.

RESULTS

A total of 31 blood-entry components were identified, corresponding to 255 potential targets associated with LOH. Metabolomics analysis revealed 48 differential metabolites, mainly involved in metabolic pathways such as glycerophospholipid metabolism, bile acid metabolism, and autophagy. Integrative analysis focused on the PI3K/Akt/mTOR signaling pathway and four core targets: AR, mTOR, GSK3β, and APP. In vivo experiments demonstrated that XCYSF restored testosterone levels in LOH rats and exerted therapeutic effects by inhibiting the PI3K/Akt/mTOR pathway. In vitro, XCYSF-containing serum and its potential key component, tribuloside, promoted testosterone secretion through inhibition of the same pathway.

CONCLUSIONS

XCYSF and its active component tribuloside promote testosterone synthesis and alleviate LOH by inhibiting the PI3K/Akt/mTOR signaling pathway.