Xie Fang, Zhou Lan, Yu Miao
Department of Traditional Chinese Medicine, The First Affiliated Hospital of Bengbu Medical University, Bengbu, 233004, China.
Department of Basic Theory of Traditional Chinese Medicine, Heilongjiang University of Chinese Medicine, Harbin, 150040, China.
Biochem Genet. 2025 Sep 12. doi: 10.1007/s10528-025-11246-5.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline and neuronal loss. Ferroptosis, a form of regulated cell death driven by iron overload and lipid peroxidation, has been implicated in AD pathology. DiHuangYinZi (DHYZ), a traditional Chinese herbal remedy, has been suggested to ameliorate cognitive impairments and reduce ferroptosis in AD models. This study aimed to investigate the effects of DHYZ on learning, memory, ferroptosis markers, and neuronal integrity in APP/PS1 transgenic mice. Six-month-old APP/PS1 transgenic mice were treated with DHYZ or donepezil for four weeks. Learning and memory functions were evaluated using the Morris Water Maze (MWM) and open field test. Neuronal integrity was assessed through Hematoxylin and Eosin (H&E) and Nissl staining. Ferroptosis markers, including superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), and the GSH/GSSG ratio, were measured in hippocampal tissues. Ferroptosis-related protein expressions, such as ferritin, DMT1, FPN1, Nrf2, and GPX4, were analyzed using Western blot. DHYZ treatment significantly improved learning and memory deficits in APP/PS1 mice, as evidenced by reduced escape latency and increased platform crossings in the MWM. DHYZ also reversed anxiety-like behavior in the open field test. Histological analysis showed that DHYZ treatment restored neuronal integrity, as indicated by better cellular arrangement and staining compared to untreated APP/PS1 mice. DHYZ inhibited ferroptosis by reducing iron overload, increasing SOD and GSH levels, and normalizing the GSH/GSSG ratio. Moreover, DHYZ modulated the expression of ferroptosis-related proteins, restoring FPN1 levels while reducing ferritin and DMT1 expressions. Nrf2 and GPX4 levels, which were reduced in APP/PS1 mice, were significantly increased after DHYZ treatment. DHYZ effectively improved cognitive deficits, inhibited ferroptosis, and restored neuronal integrity in APP/PS1 mice. These findings suggest that DHYZ may have therapeutic potential for AD by targeting ferroptosis and regulating iron metabolism.
阿尔茨海默病（AD）是一种以认知衰退和神经元丢失为特征的进行性神经退行性疾病。铁死亡是一种由铁过载和脂质过氧化驱动的程序性细胞死亡形式，已被认为与AD病理学有关。地黄饮子（DHYZ）是一种传统的中药方剂，已被证明可改善AD模型中的认知障碍并减少铁死亡。本研究旨在探讨DHYZ对APP/PS1转基因小鼠学习、记忆、铁死亡标志物和神经元完整性的影响。6月龄的APP/PS1转基因小鼠用DHYZ或多奈哌齐治疗4周。使用莫里斯水迷宫（MWM）和旷场试验评估学习和记忆功能。通过苏木精和伊红（H&E）染色及尼氏染色评估神经元完整性。在海马组织中测量铁死亡标志物，包括超氧化物歧化酶（SOD）、丙二醛（MDA）、谷胱甘肽（GSH）以及GSH/GSSG比值。使用蛋白质免疫印迹法分析铁死亡相关蛋白的表达，如铁蛋白、二价金属离子转运体1（DMT1）、铁转运蛋白1（FPN1）、核因子E2相关因子2（Nrf2）和谷胱甘肽过氧化物酶4（GPX4）。DHYZ治疗显著改善了APP/PS1小鼠的学习和记忆缺陷，MWM试验中逃避潜伏期缩短和穿越平台次数增加证明了这一点。DHYZ在旷场试验中也逆转了类似焦虑的行为。组织学分析表明，与未治疗 的APP/PS1小鼠相比，DHYZ治疗恢复了神经元完整性，表现为细胞排列和染色更好。DHYZ通过减少铁过载、增加SOD和GSH水平以及使GSH/GSSG比值正常化来抑制铁死亡。此外，DHYZ调节铁死亡相关蛋白的表达，恢复FPN1水平，同时降低铁蛋白和DMT1的表达。APP/PS1小鼠中降低的Nrf2和GPX4水平在DHYZ治疗后显著升高。DHYZ有效改善了APP/PS1小鼠的认知缺陷，抑制了铁死亡，并恢复了神经元完整性。这些发现表明，DHYZ可能通过靶向铁死亡和调节铁代谢对AD具有治疗潜力。
