Eriodictyol alleviates ovarian dysfunction in a mouse model of premature ovarian failure via the PI3K/Akt/NF-κB pathway and suppression of macrophage inflammation.

Background: Premature ovarian failure (POF) is a significant cause of female infertility characterized by amenorrhea, hypergonadotropism, and hypoestrogenism, for which effective treatments are limited. Eriodictyol, a natural flavonoid, possesses potent anti-inflammatory properties, but its effects on POF remain unexplored. This study aimed to investigate the therapeutic potential of eriodictyol in a mouse model of chemotherapy-induced POF and to elucidate its underlying molecular mechanism. Methods: A POF model was established in C57BL/6 mice by cyclophosphamide injection. Mice were then treated with eriodictyol (20, 40, or 80 mg/kg) for 4 weeks. Ovarian function was evaluated by estrous cyclicity, ovarian index, and serum hormone levels. The mechanism was investigated using a combination of computational prediction and experimental validation, including in vivo Western blotting and an in vitro macrophage-granulosa cell co-culture system. Results: Eriodictyol treatment markedly restored estrous cyclicity, increased the ovarian index, decreased serum follicle-stimulating hormone (FSH), and elevated serum estradiol (E2) and anti-Müllerian hormone (AMH) levels in POF mice. To explore the mechanism, network analysis was first employed to predict potential targets, which identified the PI3K/Akt/NF-κB signaling pathway. This computational hypothesis was then experimentally validated; Western blot analysis confirmed that eriodictyol significantly inhibited the phosphorylation of PI3K, Akt, and NF-κB p65 in ovarian tissues. Furthermore, molecular docking suggested a strong binding affinity between eriodictyol and Akt. Corroborating these findings, in vitro experiments demonstrated that eriodictyol pre-treatment of macrophages protected co-cultured granulosa cells from inflammatory damage. Conclusion: Eriodictyol alleviates chemotherapy-induced ovarian dysfunction by inhibiting the PI3K/Akt/NF-κB inflammatory pathway and suppressing macrophage-mediated damage to granulosa cells. These findings identify eriodictyol as a promising therapeutic candidate for POF.