Wang Qiang, Zuo Hanjun, Sun Huaqin, Xiao Xiao, Wang Zhao, Li Tingyu, Luo Xiaolei, Wang Yanyun, Wang Tao, Li Juanjuan, Gao Linbo
Laboratory of Molecular Translational Medicine, Center for Translational Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, China.
Department of Anatomy and Histology & Embryology, Faculty of Basic Medical Sciences, Kunming Medical University, Kunming, Yunnan, China.
CNS Neurosci Ther. 2025 Feb;31(2):e70282. doi: 10.1111/cns.70282.
Emerging evidence highlights the involvement of long non-coding RNAs (lncRNAs) and ferroptosis in the pathogenesis of traumatic brain injury (TBI). However, the regulatory role of lncRNAs in TBI-induced ferroptosis remains poorly understood. This study aims to investigate the role of a specific lncRNA, noncoding transcript of chemokine (C-C motif) ligand 4 (Ccl4) overlapping (Ntoco), in the regulation of ferroptosis following TBI and explore its potential as a therapeutic target.
The expression levels of Ntoco following controlled cortical injury (CCI) in mice were measured using real-time PCR. Behavioral tests post-injury were assessed using the rotarod test and Morris water maze, and lesion volume was evaluated using micro-MRI. Ntoco binding proteins were identified using RNA pull-down and RNA immunoprecipitation. RNA sequencing was employed to identify Ntoco-related pathways. Western blotting and co-immunoprecipitation were used to measure protein levels and ubiquitination processes.
Ntoco upregulation was observed in CCI mice. Ntoco knockdown inhibited neuron ferroptosis, reduced lesion volume, and improved spatial memory following TBI. Ntoco overexpression promoted ferroptosis in neurons. Mechanistically, Ntoco facilitated K48-linked ubiquitination and degradation of proteins by binding to Hnrnpab, suppressing the NF-κB/Lcn2 signaling pathway. This included reduced phosphorylation of IkBα, increased phosphorylation of IKKα/β, nuclear translocation of the NF-κB p65 subunit, and elevated Lcn2 expression.
Our findings suggest that Ntoco plays a crucial role in TBI-induced ferroptosis by modulating the NF-κB/Lcn2 signaling pathway. Targeting Ntoco may provide a promising therapeutic strategy to mitigate ferroptosis and improve outcomes following TBI.
新出现的证据突显了长链非编码RNA（lncRNAs）和铁死亡在创伤性脑损伤（TBI）发病机制中的作用。然而，lncRNAs在TBI诱导的铁死亡中的调控作用仍知之甚少。本研究旨在探讨一种特定的lncRNA，即趋化因子（C-C基序）配体4（Ccl4）重叠的非编码转录本（Ntoco）在TBI后铁死亡调控中的作用，并探索其作为治疗靶点的潜力。
使用实时PCR测量小鼠控制性皮质损伤（CCI）后Ntoco的表达水平。损伤后行为测试采用转棒试验和莫里斯水迷宫进行评估，损伤体积通过显微MRI进行评估。使用RNA下拉和RNA免疫沉淀鉴定Ntoco结合蛋白。采用RNA测序鉴定与Ntoco相关的通路。蛋白质印迹和免疫共沉淀用于测量蛋白质水平和泛素化过程。
在CCI小鼠中观察到Ntoco上调。Ntoco敲低抑制了神经元铁死亡，减少了损伤体积，并改善了TBI后的空间记忆。Ntoco过表达促进了神经元中的铁死亡。机制上，Ntoco通过与Hnrnpab结合促进K48连接的蛋白质泛素化和降解，抑制NF-κB/Lcn2信号通路。这包括IkBα磷酸化减少、IKKα/β磷酸化增加、NF-κB p65亚基核转位以及Lcn2表达升高。
我们的研究结果表明，Ntoco通过调节NF-κB/Lcn2信号通路在TBI诱导的铁死亡中起关键作用。靶向Ntoco可能为减轻铁死亡和改善TBI后的预后提供一种有前景的治疗策略。
