KEAP1-NRF2/HO-1通路促进精神分裂症中的铁死亡和神经元损伤。

KEAP1-NRF2/HO-1 Pathway Promotes Ferroptosis and Neuronal Injury in Schizophrenia.

作者信息

Zhu Feng, Dan Tangqun, Hua Shuguang

机构信息

Department of Psychiatry, The Second Affiliated Hospital of Hubei University of Science and Technology, Xianning, Hubei, China.

出版信息

Brain Behav. 2025 Mar;15(3):e70311. doi: 10.1002/brb3.70311.

DOI:10.1002/brb3.70311

PMID:40021790

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11870791/

Abstract

BACKGROUND

This study investigates the role of the KEAP1-NRF2/HO-1 signaling pathway in inducing ferroptosis and contributing to neuronal damage in schizophrenia.

METHODS

We retrieved schizophrenia-related data and ferroptosis-related genes from the RNA microarray dataset GSE27383 and FerrDB database, respectively. Bioinformatics data identified KEAP1 as a downregulated gene, which was validated using qRT-PCR and Western blot. We assessed intracellular Fe⁺ content, MDA levels, GSH, and GPX4 in the prefrontal cortex and peripheral blood mononuclear cells (PBMCs) of patients with schizophrenia. Cortical interneurons (cINs) were generated from human-induced pluripotent stem cells (hiPSCs) of patients with schizophrenia and used to explore KEAP1 alterations during neurodevelopment. In addition, KEAP1 overexpression was induced in cINs via transfection with pcDNA KEAP1. The intracellular Fe⁺ levels, oxidative stress indicators, lipid peroxidation, and inflammatory cytokines were measured after transfection. To investigate molecular mechanisms, KI696-a high-affinity probe that disrupts the KEAP1-NRF2 interaction-was applied, and changes in oxidative stress, lipid peroxidation (C11-BODIPY staining), iron metabolism, and inflammatory pathways were evaluated.

RESULTS

Patients with schizophrenia exhibited underexpression of KEAP1, a key regulator of ferroptosis, along with elevated intracellular Fe⁺ levels and increased MDA concentrations, indicating enhanced lipid peroxidation and oxidative stress. Reduced GPX4 activity and GSH levels were also observed, suggesting an increased susceptibility to ferroptosis. To further explore this, cINs derived from hiPSCs of patients with schizophrenia were studied. These cells showed decreased KEAP1 expression. Overexpression of KEAP1 in cINs led to a reduction in intracellular Fe⁺ concentrations and oxidative damage, highlighting KEAP1's regulatory role in ferroptosis. In addition, treatment with KI696 induced significant alterations in pathways related to oxidative stress, iron metabolism, antioxidant defenses, and inflammation.

CONCLUSION

Our findings indicate that the KEAP1-NRF2/HO-1 pathway contributes to ferroptosis and neuronal injury in schizophrenia.

摘要

背景

本研究调查KEAP1-NRF2/HO-1信号通路在精神分裂症中诱导铁死亡及导致神经元损伤方面的作用。

方法

我们分别从RNA微阵列数据集GSE27383和FerrDB数据库中检索与精神分裂症相关的数据和与铁死亡相关的基因。生物信息学数据确定KEAP1为下调基因，通过qRT-PCR和蛋白质免疫印迹法进行验证。我们评估了精神分裂症患者前额叶皮质和外周血单核细胞（PBMC）中的细胞内Fe⁺含量、丙二醛（MDA）水平、谷胱甘肽（GSH）和谷胱甘肽过氧化物酶4（GPX4）。从精神分裂症患者的人诱导多能干细胞（hiPSC）中生成皮质中间神经元（cIN），用于探索神经发育过程中KEAP1的改变。此外，通过用pcDNA KEAP1转染在cIN中诱导KEAP1过表达。转染后测量细胞内Fe⁺水平、氧化应激指标、脂质过氧化和炎性细胞因子。为了研究分子机制，应用了KI696（一种破坏KEAP1-NRF2相互作用的高亲和力探针），并评估了氧化应激、脂质过氧化（C11-硼二吡咯染色）、铁代谢和炎症途径的变化。

结果

精神分裂症患者表现出KEAP1（铁死亡的关键调节因子）表达不足，同时细胞内Fe⁺水平升高和MDA浓度增加，表明脂质过氧化和氧化应激增强。还观察到GPX4活性和GSH水平降低，表明对铁死亡的易感性增加。为了进一步探究这一点，研究了源自精神分裂症患者hiPSC的cIN。这些细胞显示KEAP1表达降低。cIN中KEAP1的过表达导致细胞内Fe⁺浓度和氧化损伤降低，突出了KEAP1在铁死亡中的调节作用。此外，用KI696处理诱导了与氧化应激、铁代谢、抗氧化防御和炎症相关途径的显著改变。