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Activation of p62-Keap1-Nrf2 Pathway Protects 6-Hydroxydopamine-Induced Ferroptosis in Dopaminergic Cells

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Abstract

Parkinson’s disease (PD) is a common neurodegenerative disorder primarily caused by the death of dopaminergic neurons in the substantia nigra pars compacta (SNpc). However, the manner of death of dopaminergic neurons remains indistinct. Ferroptosis is a form of cell death involving in the iron-dependent accumulation of glutathione depletion and lipid peroxide. Besides, previous studies indicated that ferroptosis might be involved in the death of dopaminergic neurons. In this study, we aim to explore the protective effect of the p62-Keap1-Nrf2 pathway against 6-hydroxydopamine (6-OHDA)-induced ferroptosis in dopaminergic cells. Firstly, our results demonstrated that 6-OHDA-induced ferroptosis could be observed in vivo zebrafish and in vitro human dopaminergic cell line (SH-SY5Y cells) model. Moreover, ferroptosis induced by 6-OHDA mitigates in SH-SY5Y cells upon ferrostatin-1 (Fer, an inhibitor of ferroptosis) treatment via upregulating the protein expression of glutathione peroxidase 4 (GPX4). Then, we found that high p62/SQSTM1 (p62) expression could protect SH-SY5Y cells against ferroptosis through promoting Nrf2 nuclear transfer and upregulating the expression of the antioxidant protein heme oxygenase-1 (HO-1). Ultimately, high p62 expression activates the Nrf2/HO-1 signaling pathway through binding to Kelch-like ECH-associated protein 1 (Keap1). Collectively, the activation of the p62-Keap1-Nrf2 pathway prevents 6-OHDA-induced ferroptosis in SH-SY5Y cells, targeting this pathway in combination with a pharmacological inhibitor of ferroptosis can be a potential approach for PD therapy.

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Abbreviations

PD:

Parkinson’s disease

ROS:

Reactive oxygen species

6-OHDA:

6-Hydroxydopamine

Fer:

Ferrostatin-1

HO-1:

Heme oxygenase-1

Nrf2:

Nuclear factor erythroid 2-like 2

SNpc:

Substantia nigra pars compacta

GPX4:

Glutathione peroxidase 4

ACSL4:

Acyl-CoA synthetase-4

α-syn:

α-Synuclein

Nec:

Necrosulfonamide

Keap1:

Kelch-like ECH-associated protein 1

ZnPP:

Zn-protoporphyrin

p62:

p62/SQSTM1

BCA:

Bicinchoninic acid

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Funding

This study was financially supported by the National Natural Science Foundation of China (No. 31570351) and Sichuan Province Key Research and Development Projects (2020YFS0281).

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Author notes

  1. Yiran Sun, Libo He, Taoyu Wang and Wan Hua contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, Sichuan, China

    Yiran Sun, Libo He, Taoyu Wang, Wan Hua, Huan Qin, Jingjin Wang, Li Wang, Wanqin Gu, Tingting Li, Na Li, Xinanbei Liu, Fang Chen & Lin Tang

  2. National and Local Joint Engineering Laboratory for Energy Plant Bio-Oil Production and Application, Chengdu, 610065, Sichuan, China

    Yiran Sun, Libo He, Taoyu Wang, Wan Hua, Huan Qin, Jingjin Wang, Li Wang, Wanqin Gu, Tingting Li, Na Li, Xinanbei Liu, Fang Chen & Lin Tang

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  1. Yiran Sun
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  2. Libo He
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Contributions

Yiran Sun, Libo He, Taoyu Wang, and Wan Hua designed and performed the experiments; Yiran Sun, Huan Qin, Jingjin Wang, Li Wang, Wanqin Gu, Tingting Li, Na Li, and Xinanbei Liu for formal analysis and validation; Yiran Sun wrote the original draft and Libo He, Wan Hua, and Lin Tang reviewed and edited the manuscript; Fang Chen and Lin Tang provided reagents and technical support.

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Correspondence to Lin Tang.

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All fish experiments were performed following the guidelines issued by the animal ethics committee (AAALAC Certificate NO.001458).

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Sun, Y., He, L., Wang, T. et al. Activation of p62-Keap1-Nrf2 Pathway Protects 6-Hydroxydopamine-Induced Ferroptosis in Dopaminergic Cells. Mol Neurobiol 57, 4628–4641 (2020). https://doi.org/10.1007/s12035-020-02049-3

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