Wang Xie, Shao Nan, Zhang Xiaoyan, Chen Hong, Chang Ze, Xie Daojun, Zhang Juan
The First Clinical Medical College of Anhui University of Chinese Medicine, Hefei, 230038, People's Republic of China.
Xiyuan Hospital of China Academy of Traditional Chinese Medicine, Beijing, 100091, People's Republic of China.
Neuropsychiatr Dis Treat. 2023 Dec 5;19:2681-2696. doi: 10.2147/NDT.S443278. eCollection 2023.
Wilson's disease (WD), an autosomal recessive genetic disease, is characterized by copper metabolism disorder. WD patients may have a series of cognitive deficits in terms of neurological symptoms. Ferroptosis (FPT), a type of programmed cell death, is involved in the pathological progression of various cognitive disorders, and silent information regulator 1 (SIRT1) is considered to be a key factor in FPT. Ferulic acid (FA) is a traditional Chinese medicine monomer, with a remarkable effect in the clinical treatment of cognitive impairment-related disease. However, its intrinsic effect on FPT is still unclear. This study aims to investigate the protective effect of FA on cognitive impairment in animal and cell models of WD, and whether the pharmacological mechanism is related to the SIRT1-mediated FPT signaling pathway.
Copper-loaded WD rats and PC12 cells WD were used as models of cognitive dysfunction in vivo and in vitro, respectively. Morris Water Maze (MWM) was used to evaluate the spatial exploration and memory abilities of rats. HE staining was used to observe neuronal damage in the CA1 region of the rat hippocampus. Immunofluorescence (IF) was used to detect the expression of GPX4 protein. Transmission electron microscopy (TEM) was used to observe the ultrastructure of neurons. The levels of Fe2+, MDA, SOD, GSH, 4HNE, and ROS were detected. Western blot and qRT-PCR were used to detect the protein and mRNA levels of SIRT1, Nrf2, SCL7A11, and GPX4.
In the WD copper-loaded model rats, MWM, TEM, and IF results showed that FA could promote the repair of learning and memory function, improve the morphological damage to hippocampal neurons, and maintain mitochondria integrity. In the PC12 cell experiment, the MTT method showed that FA increased the viability of copper-overloaded cell models. Western blot and qRT-PCR results confirmed that FA significantly increased the expression of proteins and mRNA in SIRT1, Nrf2, SCL7A11, and GPX4. In addition, FA reversed the expression of oxidative stress-related indicators, including MDA, SOD, GSH, 4HNE, and ROS.
FA alleviates hippocampal neuronal injury by activating SIRT1-mediated FPT, providing a valuable candidate for traditional Chinese medicine monomer for the clinical therapeutics of WD cognitive impairment.
威尔逊病（WD）是一种常染色体隐性遗传病，其特征为铜代谢紊乱。WD患者在神经症状方面可能存在一系列认知缺陷。铁死亡（FPT）是一种程序性细胞死亡，参与各种认知障碍的病理进展，而沉默信息调节因子1（SIRT1）被认为是FPT中的关键因素。阿魏酸（FA）是一种中药单体，在认知障碍相关疾病的临床治疗中具有显著效果。然而，其对FPT的内在作用仍不清楚。本研究旨在探讨FA对WD动物和细胞模型中认知障碍的保护作用，以及其药理机制是否与SIRT1介导的FPT信号通路相关。
分别以铜负荷的WD大鼠和PC12细胞作为体内和体外认知功能障碍模型。采用莫里斯水迷宫（MWM）评估大鼠的空间探索和记忆能力。采用苏木精-伊红（HE）染色观察大鼠海马CA1区的神经元损伤。采用免疫荧光（IF）检测GPX4蛋白的表达。采用透射电子显微镜（TEM）观察神经元的超微结构。检测铁离子（Fe2+）、丙二醛（MDA）、超氧化物歧化酶（SOD）、谷胱甘肽（GSH）、4-羟基壬烯醛（4HNE）和活性氧（ROS）的水平。采用蛋白质印迹法和实时定量聚合酶链反应（qRT-PCR）检测SIRT1、核因子E2相关因子2（Nrf2）、溶质载体家族7成员11（SCL7A11）和GPX4的蛋白质和mRNA水平。
在WD铜负荷模型大鼠中，MWM、TEM和IF结果显示，FA可促进学习和记忆功能的修复，改善海马神经元的形态损伤，并维持线粒体完整性。在PC12细胞实验中，MTT法显示FA增加了铜过载细胞模型的活力。蛋白质印迹法和qRT-PCR结果证实，FA显著增加了SIRT1、Nrf2、SCL7A11和GPX4中蛋白质和mRNA的表达。此外，FA逆转了氧化应激相关指标的表达，包括MDA、SOD、GSH、4HNE和ROS。
FA通过激活SIRT1介导的FPT减轻海马神经元损伤，为WD认知障碍的临床治疗提供了一种有价值的中药单体候选药物。
