The Bushen Huatan formula inhibits TP53-Mediated ferroptosis and modulating lipid metabolism in polycystic ovary syndrome abnormalities rat model.

Abstract

BACKGROUND: Bushen Huatan Formula (BSHT) shows promise in treating reproductive endocrine disorders, yet its mechanism in polycystic ovary syndrome (PCOS) remains unclear. This study aims to elucidate the specific mechanism of action of BSHT in treating PCOS. METHODS: PCOS model in rats was established by using a high-fat diet combined with letrozole gavage, and an in vitro model was constructed by stimulating human ovarian granulosa cells with dehydroepiandrosterone. Oestrus cycle changes were monitored throughout the BSHT intervention period via daily vaginal smear. Following intervention, ovarian histopathology and serum sex hormones, lipid profiles, oxidative stress, and ferroptosis markers were assessed. Network pharmacology predicted BSHT targets and pathways, validated in vitro/in vivo via TP53 pathway expression and downstream factors, oxidative stress, and key ferroptosis protein levels. RESULTS: High-dose BSHT effectively improved estrous cycle irregularities, reduced ovarian and uterine indexes, and repaired ovarian pathology in PCOS rats. BSHT treatment significantly improved endocrine profiles by reducing testosterone and luteinizing hormone levels while increasing follicle-stimulating hormone (P&#x2009;<&#x2009;0.001), and ameliorated dyslipidemia by lowering total cholesterol and low-density lipoprotein cholesterol while elevating high-density lipoprotein cholesterol (P&#x2009;<&#x2009;0.001). Mechanistically, BSHT suppressed oxidative stress and ferroptosis, as evidenced by decreased malondialdehyde, reactive oxygen species, lipid peroxides, Fe2&#x2009;+&#x2009;and ACSL4, along with increased GSH and GPX4 (P&#x2009;<&#x2009;0.001). Both network pharmacology and experimental validation confirmed that these effects were mediated through inhibition of the TP53 signaling pathway and its downstream targets (P&#x2009;<&#x2009;0.001). The critical role of TP53 was further verified through intervention studies using ferroptosis inhibitors and TP53 agonists (P&#x2009;<&#x2009;0.001). CONCLUSION: This study demonstrates that BSHT alleviates PCOS pathology by inhibiting ferroptosis and regulating lipid metabolism via the TP53 signaling pathway, providing a theoretical basis for novel therapeutic strategies.