Department of Anesthesiology, East Hospital, Renmin Hospital of Wuhan University, No. 17 Gaoxin 6th Road, Wuhan, 430223, Hubei, China.
Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei, China.
Neuromolecular Med. 2024 Nov 7;26(1):44. doi: 10.1007/s12017-024-08813-1.
This study aimed to investigate the role of ubiquitin-specific peptidase 15 (USP15) in ischemic cognitive dysfunction using a mouse model and a cerebral ischemia (CI) cell model, its impact on ferroptosis and the underlying mechanisms. Oxygen-glucose deprivation/reoxygenation (OGD/ R)-induced HT-22 cells were used to establish the CI cell model, and mice induced with CI were used as the animal model for ischemic cognitive dysfunction. Cell damage was evaluated using Cell Counting Kit-8 (CCK-8), flow cytometry (FCM), immunoblotting, and immunofluorescence assays. Cognitive dysfunction in the CI mice was assessed through water maze experiments. Ferroptosis was examined with an iron detection kit and immunoblotting, oxidative stress was evaluated using 2',7'-dichlorofluorescin diacetate (DCF) and enzyme-linked immunosorbent assay (ELISA), and mechanistic experiments were performed via immunoblotting. USP15 knockdown alleviated OGD/ R-induced damage in HT-22 cells. In vivo, USP15 depletion mitigated brain injury in middle cerebral artery occlusion (MCAO) mice and improved learning and memory function. The absence of USP15 reduced oxidative stress in MCAO mice and attenuated ferroptosis by activating nuclear factor erythroid 2-related factor 2 (Nrf2). Mechanistic investigations confirmed that USP15 depletion ameliorated cognitive impairment and ferroptosis through the activation of the Nrf2/ GPX4 axis. USP15 is associated with ferroptosis and cognitive dysfunction in mice and could serve as a potential therapeutic target in CI.
本研究旨在通过建立小鼠模型和脑缺血（CI）细胞模型，研究泛素特异性肽酶 15（USP15）在缺血性认知功能障碍中的作用，及其对铁死亡的影响和潜在机制。采用氧葡萄糖剥夺/复氧（OGD/R）诱导 HT-22 细胞建立 CI 细胞模型，采用脑缺血诱导的小鼠作为缺血性认知功能障碍的动物模型。采用细胞计数试剂盒-8（CCK-8）、流式细胞术（FCM）、免疫印迹和免疫荧光法评估细胞损伤。通过水迷宫实验评估 CI 小鼠的认知功能障碍。使用铁检测试剂盒和免疫印迹法检测铁死亡，使用 2'，7'-二氯荧光素二乙酸酯（DCF）和酶联免疫吸附试验（ELISA）评估氧化应激，通过免疫印迹法进行机制实验。USP15 敲低减轻了 HT-22 细胞 OGD/R 诱导的损伤。在体内，USP15 耗竭减轻了大脑中动脉闭塞（MCAO）小鼠的脑损伤并改善了学习和记忆功能。USP15 缺失减少了 MCAO 小鼠的氧化应激并通过激活核因子红细胞 2 相关因子 2（Nrf2）减轻铁死亡。机制研究证实，USP15 耗竭通过激活 Nrf2/GPX4 轴改善认知障碍和铁死亡。USP15 与小鼠的铁死亡和认知功能障碍有关，可能成为 CI 的潜在治疗靶点。
