The effect of norepinephrine on ovarian dysfunction by mediating ferroptosis in mice model

Studies shows that stress is associated with ovarian dysfunction. Norepinephrine (NE), a classic stress hormone in the stress response, is less recognized for its role in ovarian function. A NE-treated mouse model is induced by intraperitoneal injection of NE for 4 weeks. Compared with the normal control, we find that NE-treated mice show disturbances in the estrous cycle, decreased levels of anti-Mullerian hormone (AMH) and estradiol (E2), and increased levels of follicle-stimulating hormone (FSH). Additionally, the number of primordial follicles, primary follicles, secondary follicles, and antral follicles decreased, while the number of atretic follicles increased in NE-treated mice, indicating NE-induced ovarian dysfunction. RNA sequencing further reveals that genes associated with ferroptosis are significantly enriched in NE-treated ovarian tissues. Concurrently, the content of reactive oxygen species (ROS), ferrous ion, and malondialdehyde (MDA) increased, while the expression level of glutathione peroxidase 4 (GPX4) decreased.

To elucidate the mechanism of NE-induced ferroptosis in ovaries and the potential reversal by Coenzyme Q10 (CoQ10), an antioxidant, we conduct both in vitro and in vivo experiments. In vitro, we observe that the granulosa cell line KGN, when treated with NE, shows decreased cell viability, reduced expression of GPX4, elevated ferrous ion and ROS content, and increased MDA levels. However, these NE-induced changes are rescued by the addition of CoQ10. In the mouse model, we find that NE-treated mice supplemented with CoQ10 increased GPX4 levels and decreased the contents of iron, ROS, and MDA compared with the NE group. Moreover, the differential expression of genes associated with ferroptosis induced by NE is ameliorated by CoQ10 in NE-treated mice. Additionally, CoQ10 improved ovarian function, as evidenced by increased ovarian weight, more regular estrous cycles, and an increase in follicles at various stages of growth in NE-treated mice. In conclusion, NE induces ovarian dysfunction by triggering ferroptosis in ovarian tissues, and CoQ10 represents a promising approach for protecting reproductive function by inhibiting ferroptosis.