Honokiol alleviates radiation‐induced premature ovarian failure via enhancing Nrf2

Abstract Background The ovary is highly sensitive to radiation, and patients receiving radiotherapy are at significant risk of premature ovarian failure (POF). This study aimed to explore the radioprotective effect of honokiol (HKL) on ionizing radiation (IR)‐induced POF. Methods Female C57BL/6 mice were administered intraperitoneally with vehicle or HKL once daily for 7 days. On day 7, the mice in the IR and HKL+IR groups were exposed to 3.2 Gy whole‐body radiation for one hour after the intraperitoneal injection and sacrificed 12 or 72 h after radiation exposure. The gonadosomatic index (GSI) was calculated. Blood samples were collected for enzyme‐linked immunosorbent assay (ELISA). Ovaries were harvested for histological examination, immunohistochemistry, immunofluorescence, TUNEL, western blot, and qPCR. The fertility assessment was evaluated by calculating live offspring number. Results The optimum dose of HKL against radiation was 10 mg/kg via intraperitoneal injection. POF was induced 72 h after irradiation with significantly downregulated proliferating cell nuclear antigen (PCNA). The numbers of primordial and preantral follicles decreased significantly after irradiation ( p < .001), whereas the number of atretic follicles increased ( p < .001). The serum levels of estradiol (E 2 ) and anti‐Müllerian hormone (AMH) decreased to 50% of the control group after irradiation ( p < .05). Moreover, the GSI after irradiation was 27% lower than in the control group ( p < .05). The number of offspring in the IR group dropped by 50% compared with the control group ( p < .05). HKL pretreatment protected the animals’ fertility, GSI, PCNA, serum levels of E 2 and AMH, and the number of primordial and preantral follicles. Significant upregulation of apoptosis‐related proteins such as Pho‐P53, Bax, Cyto C, C‐caspase‐3, C‐PARP, and pyroptosis‐related proteins such as Pho‐NF‐κB p65, NLRP3, caspase‐1, IL‐1β, and IL‐18 was observed after irradiation, while the expression of Bcl‐2 decreased. HKL pretreatment prevented these changes. After irradiation, malondialdehyde (MDA), Nrf2, and HO‐1 were upregulated. HKL treatment activated the expression of Nrf2 and HO‐1 and promoted the nucleus translocation of Nrf2. Furthermore, HKL did not affect ovarian reserves under physiological conditions. Conclusions HKL ameliorated IR‐induced POF by inhibiting apoptosis and pyroptosis by enhancing Nrf2 expression and translocation.