Fang Jiang, Yuan Qiang, Du Zhuoying, Fei Maoxing, Zhang Quan, Yang Lei, Wang Meihua, Yang Weijian, Yu Jian, Wu Gang, Hu Jin
Department of Neurosurgery, Huashan Hospital, Fudan University, Neurosurgical Institute of Fudan University, Shanghai Clinical Medical Center of Neurosurgery, Shanghai Key Laboratory of Brain Function Restoration and Neural Regeneration, Shanghai, China.
Department of Neurosurgery, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.
Biochem Biophys Res Commun. 2022 Sep 3;619:34-41. doi: 10.1016/j.bbrc.2022.06.040. Epub 2022 Jun 14.
Ferroptosis is a newly recognized form of regulated cell death. Recently, growing evidence has shown that ferroptosis is involved in the pathogenesis of traumatic brain injury (TBI). However, less attention has been paid to its role in brain microvascular endothelial cells (BMVECs) and blood-brain barrier (BBB) damage, the central pathological process in secondary brain injury of TBI. Here, we established a mechanical stretch injury bEnd.3 model and a Controlled Cortical Impact (CCI) mouse model to explore the ferroptosis-related markers in brain endothelial cells after TBI in vitro and in vivo. From the results of RNA-seq analysis, RT-qPCR and immunostaining, glutathione peroxidase 4 (GPX4) downregulation, Cyclooxygenase-2 (COX-2) upregulation, and iron accumulation were observed in brain endothelial cells after TBI both in vitro and in vivo. Furthermore, we utilized Ferrostatin-1 (Fer-1), a specific inhibitor of ferroptosis, to investigate the protective effects of ferroptosis inhibition on BBB disruption and neurological deficits. From the results of immunostaining, transmission electron microscopy (TEM), and western blotting, we demonstrated that Fer-1 significantly reduced BMVECs death, BBB permeability, and tight junction loss at 3 days after TBI. The neurological tests including grid walking, rotarod test, and wire-hanging test showed that Fer-1 administration exerted neuroprotective effects in the early stage of TBI. Our findings provided evidences for inhibition of BMVECs ferroptosis as a promising therapeutic target against TBI-induced BBB disruption.
铁死亡是一种新发现的程序性细胞死亡形式。最近，越来越多的证据表明铁死亡参与了创伤性脑损伤（TBI）的发病机制。然而，其在脑微血管内皮细胞（BMVECs）和血脑屏障（BBB）损伤中的作用却较少受到关注，而这正是TBI继发性脑损伤的核心病理过程。在此，我们建立了机械拉伸损伤bEnd.3模型和控制性皮质撞击（CCI）小鼠模型，以探究TBI后体外和体内脑内皮细胞中铁死亡相关标志物。通过RNA测序分析、逆转录定量聚合酶链反应（RT-qPCR）和免疫染色结果，发现在TBI后的体外和体内脑内皮细胞中均观察到谷胱甘肽过氧化物酶4（GPX4）下调、环氧化酶-2（COX-2）上调以及铁蓄积。此外，我们使用铁死亡特异性抑制剂铁抑素-1（Fer-1）来研究抑制铁死亡对BBB破坏和神经功能缺损的保护作用。通过免疫染色、透射电子显微镜（TEM）和蛋白质印迹法结果表明，Fer-1在TBI后3天显著减少了BMVECs死亡、BBB通透性和紧密连接丧失。包括网格行走、转棒试验和悬线试验在内的神经学测试表明，给予Fer-1在TBI早期发挥了神经保护作用。我们的研究结果为抑制BMVECs铁死亡作为对抗TBI诱导的BBB破坏的一个有前景的治疗靶点提供了证据。
