Ma Jialiang, Wang Hongxia, Jia Juan, Tao Ting, Shan Lingzhi, Sun Shougang, Wang Manxia
Lanzhou University Second Hospital, Lanzhou, 730030, China.
Gansu University of Chinese Medicine, Lanzhou, 730030, China.
Mol Neurobiol. 2025 Mar 5. doi: 10.1007/s12035-025-04800-0.
The impact of empagliflozin on OGD/R-induced ferroptosis in neurons is still unclear. This study aims to explore whether ferroptosis is associated with OGD/R-induced neuronal injury and the effect of empagliflozin on the ferroptosis effect of OGD/R-treated neurons. Western blotting, immunofluorescence, and RT-qPCR were used to detect the protein and mRNA levels of GPX4, Nrf2, and HO-1. ELISA, flow cytometry, and confocal microscopy were applied to analyze oxidative stress. Transmission electron microscopy and CCK-8 were used to determine the degree of ferroptosis in neurons. We observed a reduction in GPX4 levels and an increase in Nrf2 and HO-1 levels in OGD/R related neurons HT-22 cells. Notably, OGD/R elevates lipid peroxidation accumulation, ROS, Fe, and MDA levels while reducing GSH levels and decreasing mitochondrial membrane potential, leading to abnormal mitochondrial structure and eventual neuronal ferroptosis. Empagliflozin activates the Nrf2/HO-1 signaling pathway, enhances cellular antioxidant capacity, inhibits lipid peroxidation in OGD/R-treated neurons, and restores cellular iron homeostasis. In addition, empagliflozin can significantly reverse ferroptosis in OGD/R-treated neurons, and overexpression of Nrf2 combined with empagliflozin further inhibits ferroptosis in OGD/R-treated neurons. These results suggest that ferroptosis may be an essential cause of OGD/R-related neuron death. Empagliflozin exhibits a protective influence against OGD/R-induced ferroptosis by activating the Nrf2/HO-1 pathway.
恩格列净对氧糖剥夺/复氧（OGD/R）诱导的神经元铁死亡的影响仍不清楚。本研究旨在探讨铁死亡是否与OGD/R诱导的神经元损伤相关，以及恩格列净对OGD/R处理的神经元铁死亡效应的影响。采用蛋白质免疫印迹法、免疫荧光法和逆转录-定量聚合酶链反应（RT-qPCR）检测谷胱甘肽过氧化物酶4（GPX4）、核因子E2相关因子2（Nrf2）和血红素氧合酶-1（HO-1）的蛋白质和mRNA水平。应用酶联免疫吸附测定（ELISA）、流式细胞术和共聚焦显微镜分析氧化应激。采用透射电子显微镜和细胞计数试剂盒-8（CCK-8）测定神经元的铁死亡程度。我们观察到在OGD/R处理的神经元HT-22细胞中，GPX4水平降低，Nrf2和HO-1水平升高。值得注意的是，OGD/R会增加脂质过氧化积累、活性氧（ROS）、铁和丙二醛（MDA）水平，同时降低谷胱甘肽（GSH）水平并降低线粒体膜电位，导致线粒体结构异常并最终导致神经元铁死亡。恩格列净激活Nrf2/HO-1信号通路，增强细胞抗氧化能力，抑制OGD/R处理的神经元中的脂质过氧化，并恢复细胞铁稳态。此外，恩格列净可显著逆转OGD/R处理的神经元中的铁死亡，Nrf2过表达与恩格列净联合使用可进一步抑制OGD/R处理的神经元中的铁死亡。这些结果表明，铁死亡可能是OGD/R相关神经元死亡的重要原因。恩格列净通过激活Nrf2/HO-1途径对OGD/R诱导的铁死亡具有保护作用。
