USP2通过稳定SK-N-SH细胞中的FOXC1来减轻MPP+诱导的神经元损伤。

IF 2.7 4区医学 Q3 NEUROSCIENCES

Brain Research Pub Date : 2025-05-10 DOI:10.1016/j.brainres.2025.149689

Zhijiang Zhuang , Lihong Zhang , Yizhao Wang

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引用次数: 0

摘要

背景：叉头盒转录因子在各种器官系统的发育中起着至关重要的作用，并在包括帕金森病（PD）在内的许多神经退行性疾病中表现出神经保护作用。然而，叉头盒C1 （FOXC1）在PD发病机制中的作用和机制尚不明确。方法：用1-甲基-4-苯基吡啶（MPP+）处理人神经母细胞瘤SK-N-SH细胞，建立PD体外模型。采用实时定量聚合酶链反应（RT-qPCR）检测FOXC1和泛素特异性肽酶2 (USP2) mRNA水平。Western blot检测FOXC1、b细胞淋巴瘤-2 （Bcl-2）、Bcl-2相关 X 蛋白（Bax）、Cleaved caspase-3和USP2蛋白水平。采用CCK-8法和流式细胞术分析细胞活力和凋亡情况。ELISA法检测肿瘤坏死因子α (TNF-α)、白细胞介素-1β (IL-1β)、IL-6水平。采用专用检测试剂盒检测活性氧（ROS）、丙二醛（MDA）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽（GSH）产物。在Ubibrowser在线数据库预测后，使用共免疫沉淀（CoIP）法验证USP2与FOXC1之间的相互作用。结果：PD患者和MPP+处理的SK-N-SH细胞FOXC1和USP2表达降低。MPP+处理可引起体外SK-N-SH细胞活力抑制、炎症反应、氧化应激和细胞凋亡促进。此外，过表达FOXC1可减轻体外MPP+诱导的SK-N-SH细胞损伤。机制上，USP2直接与FOXC1相互作用，使FOXC1去泛素化，从而增强FOXC1蛋白的稳定性。结论：USP2通过稳定FOXC1减轻MPP+引发的SK-N-SH细胞损伤，为PD治疗提供了一个有希望的治疗靶点。

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USP2 alleviates MPP+-induced neuronal injury by stabilizing FOXC1 in SK-N-SH cells

Background

Forkhead box transcription factors play a crucial role in the development of various organ systems, and exhibit neuroprotective properties in many neurodegenerative diseases, including Parkinson’s disease (PD). However, the role and mechanism of Forkhead box C1 (FOXC1) in the pathogenesis of PD is poorly defined.

Methods

Human neuroblastoma SK-N-SH cells were treated with 1-methyl-4-phenylpyridinium (MPP⁺) to establish an in vitro model of PD. FOXC1 and Ubiquitin-specific peptidase 2 (USP2) mRNA levels were detected using real-time quantitative polymerase chain reaction (RT-qPCR). FOXC1, B-cell lymphoma-2 (Bcl-2), Bcl-2 related X protein (Bax), Cleaved caspase-3, and USP2 protein levels were determined using Western blot. Cell viability and apoptosis were analyzed using CCK-8 assay and flow cytometry. Tumor necrosis factor α (TNF-α), Interleukin-1β (IL-1β), and IL-6 levels were analyzed using ELISA. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and Glutathione (GSH) products were examined using special assay kits. After Ubibrowser online database prediction, the interaction between USP2 and FOXC1 was verified using Co-immunoprecipitation (CoIP) assay.

Results

FOXC1 and USP2 expression were decreased in PD patients and MPP⁺-treated SK-N-SH cells. MPP⁺ treatment could elicit SK-N-SH cell viability inhibition, inflammatory response, oxidative stress, and apoptosis promotion in vitro. Furthermore, overexpressing FOXC1 relieved MPP⁺-induced SK-N-SH cell injury in vitro. Mechanistically, USP2 directly interacted with FOXC1 and deubiquitinated FOXC1, therefore enhancing FOXC1 protein stability.

Conclusion

USP2 attenuated MPP⁺-triggered SK-N-SH cell injury through stabilizing FOXC1, providing a promising therapeutic target for PD treatment.

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来源期刊

Brain Research 医学-神经科学

CiteScore

5.90

自引率

3.40%

发文量

268

审稿时长

47 days

期刊介绍： An international multidisciplinary journal devoted to fundamental research in the brain sciences. Brain Research publishes papers reporting interdisciplinary investigations of nervous system structure and function that are of general interest to the international community of neuroscientists. As is evident from the journals name, its scope is broad, ranging from cellular and molecular studies through systems neuroscience, cognition and disease. Invited reviews are also published; suggestions for and inquiries about potential reviews are welcomed. With the appearance of the final issue of the 2011 subscription, Vol. 67/1-2 (24 June 2011), Brain Research Reviews has ceased publication as a distinct journal separate from Brain Research. Review articles accepted for Brain Research are now published in that journal.