Lianyungang Hospital of Traditional Chinese Medicine, 222004, Lianyungang, China.
School of Chinese Medicine, School of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, 210023, Nanjing, China.
J Ethnopharmacol. 2024 Jan 30;319(Pt 2):117226. doi: 10.1016/j.jep.2023.117226. Epub 2023 Sep 23.
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 Fe, 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 Fe, 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.
中风后抑郁症（PSD）是一种在中风后表现出明显的情绪低落和兴趣减退的病症。地黄饮子（DHYZ）是一种传统的中药制剂，在 PSD 治疗中得到了广泛的应用，并显示出良好的效果。然而，该配方对 PSD 的联合作用机制尚不清楚。
本研究旨在评估 DHYZ 提取物对 PSD 大鼠的治疗效果，并进一步探讨其潜在机制。
采用高效液相色谱-紫外法（HPLC-UV）测定 DHYZ 提取物的活性成分。建立 PSD 大鼠模型后，进行神经功能和抑郁样行为测试，评估 DHYZ 提取物的神经保护作用。采用 TTC 染色评估脑组织损伤体积，采用透射电镜（TEM）观察前额叶皮层区神经元的超微结构变化，采用 TUNEL 法评估大脑前额叶皮层区的细胞凋亡。采用 TTC 染色评估脑组织中铁、丙二醛（MDA）、活性氧（ROS）和谷胱甘肽（GSH）等铁死亡标志物的变化。此外，采用 Western blot 或 PCR 分别检测 P53/SLC7A11 信号通路关键蛋白或 mRNA 水平的表达。另外，还构建了 P53 过表达载体，以增加 P53 的水平。在这一系列实验中，评估了铁死亡标志物和 P53/SLC7A11 信号通路的关键因素。
DHYZ 提取物可增加 SPT 中的蔗糖偏好，但减少 PSD 大鼠 FST 的不动时间和皮质梗死体积。TEM 研究显示，DHYZ 提取物改善了 PSD 大鼠皮质区的突触超微结构。此外，DHYZ 治疗有效降低了 ROS 和 MDA 水平，抑制了铁死亡相关标志物如 Fe、SLC7A11 和 GPX4 的表达。此外，DHYZ 促进了 P53 的泛素化，从而抑制了其降解。值得注意的是，AAV 介导的 P53 过表达逆转了 DHYZ 对 PSD 大鼠神经保护和铁死亡抑制的作用。
我们的研究结果表明，DHYZ 提取物可缓解 PSD 大鼠的症状并增强其功能能力，主要通过抑制 P53/SLC7A11/GPX4 通路的铁死亡来实现。
