Acupuncture modulates extracellular vesicle-miRNA-HMOX1 axis to inhibit ferroptosis in polycystic ovary syndrome: integrating machine learning, in vitro and in vivo studies.

BACKGROUND: Acupuncture therapy shows promise for polycystic ovary syndrome (PCOS), but its molecular mechanisms remain unclear. This study investigates whether acupuncture alleviates PCOS by regulating granulosa cell (GC) ferroptosis via extracellular vesicles (EVs). METHODS: Bioinformatic analysis of KGN cell transcriptome data identified heme oxygenase-1 (HMOX1) as a key candidate. Its role was validated in a PCOS rat model and GCs using RT-qPCR, western blot, and immunohistochemistry. Ferroptosis was assessed via malondialdehyde (MDA), glutathione (GSH), and ferroptosis-related proteins (GPX4, FTH1, SLC7A11, NCOA4). HMOX1 gain/loss-of-function, rescue experiments (ferrostatin-1), and transmission electron microscopy defined its function. PCOS rats received acupuncture for 4 weeks; ovarian function and ferroptosis markers were analyzed. EVs from follicular fluid were characterized, and miRNA candidates targeting HMOX1 were screened. The EV-miRNA mechanism was tested via co-culture and inhibition assays. RESULTS: HMOX1 was upregulated in PCOS. Its overexpression induced ferroptosis and impaired GC proliferation, reversible by ferrostatin-1. Acupuncture improved ovarian histology and oocyte maturation, concurrently normalizing HMOX1 and ferroptosis markers. It specifically enriched miR-873-5p in EVs. These EVs delivered miR-873-5p to GCs, downregulating HMOX1 and alleviating ferroptosis, an effect blocked by miR-873-5p inhibition. CONCLUSIONS: Acupuncture mitigates PCOS by promoting EV-mediated transfer of miR-873-5p, which targets HMOX1 to inhibit GC ferroptosis and restore ovarian function. This identifies a novel acupuncture-EV-miRNA-HMOX1-ferroptosis axis as a therapeutic mechanism for PCOS.