Di-Huang-Yin-Zi regulates P53/SLC7A11 signaling pathway to improve the mechanism of post-stroke depression

Post-stroke depression (PSD) is a condition characterized by a profoundly depressed mood and diminished interest following a stroke. Di-Huang-Yin-Zi (DHYZ), a traditional Chinese herbal preparation, gained widespread use and shown favorable outcomes in PSD treatment. However, the combination mechanisms of this formula for PSD remain unclear.This study aimed to assess the therapeutic effects of DHYZ extract on rats with PSD and further investigate its underlying mechanism.The active components of DHYZ extract were quantified by the high-performance liquid chromatography-ultraviolet (HPLC-UV). Neurofunctional and depressive-like behavioral tests were performed to assess the neuroprotective effects of DHYZ extract after establishing a PSD rat model. Brain tissue damage volume was assessed using TTC staining, and transmission electron microscopy (TEM) was used to observe the ultrastructural changes of neurons in the prefrontal cortex region, while cell apoptosis was evaluated through TUNEL assay in the prefrontal cortex region of the brain. The effect of DHYZ on ferroptosis markers includes Fe2+, malondialdehyde (MDA), reactive oxygen species (ROS), and glutathione (GSH) were determined in the brain tissue. Moreover, the expression of key proteins or mRNA levels of the P53/SLC7A11 signaling pathway were detected using Western blot or PCR, respectively. Additionally, P53-constructed overexpression vectors were injected to increase the level of P53. In this series of experiments, ferroptosis markers and key factors of the P53/SLC7A11 signaling pathway were evaluated.DHYZ extract could increase the sucrose preference of SPT, but decrease the duration of immobility of FST and cortical infarct volume of PSD rats. A TEM study revealed that DHYZ extract improved synaptic ultrastructure in the cortical region of PSD rats. Furthermore, DHYZ treatment effectively decreased ROS and MDA levels, inhibiting the expression of ferroptosis-related markers such as Fe2+, SLC7A11, and GPX4. Additionally, DHYZ promoted the ubiquitination of P53, thus inhibiting its degradation. Notably, AAV-mediated overexpression of P53 reversed the effects of DHYZ on neuroprotection and ferroptosis inhibition in PSD rats.Our results demonstrated that DHYZ extract alleviates the symptoms and enhances the functional capability of PSD rats, mainly by suppressing the ferroptosis through the P53/SLC7A11/GPX4 pathway.