Zhao Qiuju, Wang Xinya, Zhang Taiyu, Guo Shuxian, Liu Xiaojia, Wan Shihao, Guo Yani, Zhang Qiannuo, Li Jinshui, Wang Shaohua, Dong Liuyi
Department of Pharmacology, Key Laboratory of Anti-Inflammatory and Immunopharmacology of Ministry of Education, Key Laboratory of Chinese Medicine Research and Development of State Administration of Traditional Chinese Medicine, Anhui Medical University, Hefei, Anhui, PR China.
Department of Neurosurgery, Hefei Hospital Affiliated to Anhui Medical University, The Second People's Hospital of Hefei, Hefei, Anhui, PR China.
Food Chem Toxicol. 2025 Mar;197:115281. doi: 10.1016/j.fct.2025.115281. Epub 2025 Jan 27.
Ischemic stroke is a very common brain disorder. This study aims to assess the neuroprotective effects of piceatannol (PCT) in preventing neuronal injury resulting from cerebral ischemia and reperfusion (I/R) in mice. Additionally, we investigated the underlying mechanisms through which PCT inhibits neuronal ferroptosis by modulating the USP14/GPX4 signaling axis. In vitro and in vivo experiments were conducted. In vitro, oxygen-glucose deprivation followed by reoxygenation (OGD/R) was used to simulate ischemic injury in neuronal cells. We utilized various techniques, including DCFH-DA staining, FeRhoNox-1 staining, MDA and GSH determination, immunofluorescence, Western blotting, co-immunoprecipitation, plasmid and siRNA transfection, to evaluate the therapeutic efficacy of PCT and elucidate its mechanism of action. For vivo studies, we established a mouse model of I/R by ligating the bilateral common carotid arteries. The efficacy of PCT in mitigating brain injury and cognitive dysfunction were assessed through behavioral tests, histological analysis, Western blotting, and immunohistochemistry. PCT treatment significantly enhanced cell viability under OGD/R and reduced lipid peroxidation by decreasing levels of ROS, MDA. Furthermore, PCT effectively inhibited neuronal ferroptosis by modulating the expression of key ferroptosis-related proteins, including GPX4, ACSL4, FPN1, and Ferritin. Mechanistically, PCT was found to prevent GPX4 degradation through USP14-mediated deubiquitination. Notably, silencing USP14 reversed the ferroptotic effects of PCT, whereas overexpressing of USP14 amplified these effects. In vivo, PCT significantly reduced pathological damage of brain tissue and improved cognitive dysfunction. Piceatannol exerts neuroprotective effects by regulating ferroptosis through the USP14/GPX4 axis, thereby preventing cerebral ischemia/reperfusion injury.
缺血性中风是一种非常常见的脑部疾病。本研究旨在评估白藜芦醇（PCT）在预防小鼠脑缺血再灌注（I/R）所致神经元损伤方面的神经保护作用。此外，我们还研究了PCT通过调节USP14/GPX4信号轴抑制神经元铁死亡的潜在机制。进行了体外和体内实验。在体外，采用氧糖剥夺后再给氧（OGD/R）来模拟神经元细胞的缺血性损伤。我们运用了多种技术，包括DCFH-DA染色、FeRhoNox-1染色、丙二醛（MDA）和谷胱甘肽（GSH）测定、免疫荧光、蛋白质免疫印迹法、免疫共沉淀、质粒和小干扰RNA（siRNA）转染，以评估PCT的治疗效果并阐明其作用机制。对于体内研究，我们通过结扎双侧颈总动脉建立了I/R小鼠模型。通过行为测试、组织学分析、蛋白质免疫印迹法和免疫组织化学评估了PCT减轻脑损伤和认知功能障碍的效果。PCT处理显著提高了OGD/R条件下的细胞活力，并通过降低活性氧（ROS）、MDA水平减少了脂质过氧化。此外，PCT通过调节关键铁死亡相关蛋白（包括GPX4、ACSL4、FPN1和铁蛋白）的表达有效抑制了神经元铁死亡。从机制上讲，发现PCT通过USP14介导的去泛素化作用防止GPX4降解。值得注意的是，沉默USP14可逆转PCT的铁死亡效应，而过度表达USP14则会放大这些效应。在体内，PCT显著减轻了脑组织的病理损伤并改善了认知功能障碍。白藜芦醇通过USP14/GPX4轴调节铁死亡发挥神经保护作用，从而预防脑缺血/再灌注损伤。
