Bushen Shugan Huayu formula improves depressive-like behavior in perimenopausal mice by modulating neuronal ferroptosis through the GPER1/Nrf2 pathway.

BACKGROUND

Perimenopausal depression (PMD) is closely associated with estrogen deficiency, but current therapeutic strategies for PMD have limited effectiveness and severe side effects. Accumulating evidence underscores the critical role of ferroptosis in the pathogenesis of depression.

OBJECTIVE

This research aimed to evaluate the therapeutic efficacy of Bushen Shugan Huayu formula (BSSGHY) in alleviating the symptoms of PMD and elucidate whether the underlying mechanism involves inhibition of neuronal ferroptosis through modulating the GPER1/Nrf2 signaling pathway.

METHODS

The active constituents of BSSGHY were identified using UPLC-MS/MS analysis. Behavioral analyses, histological analyses, immunoblotting, and transmission electron microscopy observations were performed in vivo in a perimenopausal mouse model (n = 10 per group). Key targets were subsequently identified through network pharmacology, validated using molecular docking simulation, and Cellular Thermal Shift Assay (CETSA). In addition, in vitro validation was conducted using erastin-induced ferroptosis and estrogen receptor-related gene knockdown in neuronal cells.

RESULTS

BSSGHY ameliorated depressive-like behaviors and hormonal imbalance in OVX mice (p < 0.05). It improved mitochondrial morphology, restored hippocampal synaptic plasticity, and attenuated ferroptosis by reducing lipid peroxidation and modulating ferroptosis-related protein expression (p < 0.05). Network pharmacology, molecular docking analysis, and CETSA have shown that multiple active compounds present in BSSGHY have high binding affinity for GPER1. In vitro experiments confirmed that BSSGHY activated the GPER1/Nrf2 axis, whereas GPER1 knockdown attenuated these protective effects.

CONCLUSION

Our study is the first, to the best of our knowledge, to demonstrate that BSSGHY exhibits antidepressant properties in PMD by suppressing neuronal ferroptosis via the GPER1/Nrf2 pathway. The study findings suggest that inhibiting ferroptosis could represent a promising therapeutic approach for the management of estrogen deficiency-related mood disorders.