Jinfeng Pills alleviate letrozole-induced PCOS by correcting CYP17A1/CYP19A1 steroidogenic imbalance and PI3K/Akt/mTOR-related signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Jinfeng Pills (JFP) is a classical Chinese patent medicine originating as an imperial court prescription during the Qianlong reign of the Qing dynasty (1735-1796), in continuous use in southern China and Southeast Asia for over two centuries (NMPA approval No. Z20064400). Comprising nine ingredients - Panax ginseng C.A.Mey., Cervus elaphus Linnaeus, Ligustrum lucidum W.T.Aiton, Epimedium brevicornu Maxim., Reynoutria multiflora (Thunb.) Moldenke, Leonurus japonicus Houtt., Cinnamomum cassia (L.) J.Presl, Equus asinus Linnaeus, and Curculigo orchioides Gaertn. - formulated as a honey-based pill and traditionally indicated for kidney-yang deficiency with blood stasis, its clinical presentations of oligomenorrhea and anovulatory infertility closely mirror PCOS hallmarks, yet its multi-constituent pharmacological mechanisms remain unresolved.

AIM OF THE STUDY: This study aimed to evaluate the effects of JFP in a letrozole-induced PCOS rat model and explore associated molecular pathways using network pharmacology and multi-layer molecular profiling.

MATERIALS AND

METHODS

PCOS was induced in SD rats via letrozole, followed by 21-day JFP treatment (160, 240, 320 mg/kg) or metformin. Estrous cyclicity, serum hormones, and ovarian morphology were evaluated. Network pharmacology, ovarian proteomics, qPCR, Western blot, serum metabolomics, and 16S rRNA gut microbiota profiling were performed as complementary analyses.

RESULTS

Compared with the model group, JFP at 320 mg/kg, the most effective dose among the tested doses, significantly reduced testosterone and LH, restored estradiol levels, and improved the LH/FSH ratio mainly through LH suppression, while FSH levels remained unchanged. JFP also increased corpus luteum number (P = 0.0020) and reduced atretic follicles (P = 0.0152), with no detectable hepatorenal toxicity. Network pharmacology and multi-layer molecular profiling, supported by qPCR and Western blot validation, suggested that JFP-associated improvements in reproductive endocrine disturbance and follicular developmental abnormalities were accompanied by changes in CYP17A1/CYP19A1-mediated steroidogenesis, PI3K/Akt/mTOR-related signaling, and IL-17A/COX2-and MAPK/ERK-p53-associated inflammatory-apoptotic pathways.

CONCLUSION

JFP improved endocrine and ovarian dysfunction in letrozole-induced PCOS rats, with its effects associated with modulation of CYP17A1/CYP19A1-mediated steroidogenesis, PI3K/Akt/mTOR-related signaling, and inflammatory-apoptotic pathways. These findings provide preclinical evidence and mechanistic clues for further investigation of JFP in PCOS-related reproductive endocrine disturbance and ovarian histomorphological abnormalities, while direct ovulation and fertility outcomes require further validation.