The Effect and Mechanism of Hyperin on Ovarian Reserve of Tripterygium Glycosides-Induced POI Mice

  Objective  To investigate the effect and mechanism of hyperin on the improvement of ovarian reserve of tripterygium glycosides (TG)-induced primary ovarian insufficiency (POI) in mice.

  Methods  Adult female BALB/c mice were used as research subjects and were randomly assigned to the control group, POI model group and hyperin treatment group, with 40 mice in each group. TG was given at 40 mg/kg twice a day by gavage for 2 weeks to create the POI mouse model. Mice in the hyperin treatment group were given hyperin at 75 mg/(kg·d) by gavage for 4 weeks after the model was established. The body mass of the mice was weighed and the gonadal index was calculated. Ovarian histological changes were observed by HE staining, and the number of follicles at all levels was calculated. Serum estradiol (E₂), follicle-stimulating hormone (FSH), anti-mullerian hormone (AMH), superoxide dismutase (SOD) and catalase (CAT) were assessed with ELISA. The mRNA and protein levels of nuclear factor (erythroid-derived 2)-related factor 2 (Nrf-2), heme oxygenase-1 (HO-1), Caspase3, Bcl-2 and Bax in ovarian granulosa cells were measured by RT-qPCR and Western blot. The protein levels of phosphorylated phosphatidylinositol 3-hydroxy kinase (p-PI3K) and phosphorylated protein kinase B (p-Akt) were measured by Western blot. The reactive oxygen species (ROS) levels in granulosa cells were determined by H2DCFDA. The apoptosis of granulosa cells was examined by TUNEL assay.

  Results  Compared with mice in the POI model group, the body mass and gonadal index of hyperin-treated mice increased (all P<0.05). The pathological damage of the ovary decreased, and the number of follicles at all levels and corpora lutea increased (all P<0.05). Serum E₂, AMH, SOD and CAT levels increased, and FSH level decreased (all P<0.05). At the molecular level, the expression of Nrf-2, HO-1, p-PI3K, p-Akt and Bcl-2 in ovarian granulosa cells increased, while the expression of Caspase3 and Bax decreased (all P<0.05). ROS level decreased (P<0.05). TUNEL assay showed reduced apoptosis of granulosa cells (P<0.05).

  Conclusion  Hyperin improved ovarian reserve in TG-induced POI mice through Nrf-2/HO-1antioxidant stress response and the anti-apoptotic effect of PI3K/Akt pathways.