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Artemisinin inhibits neuronal ferroptosis in Alzheimer’s disease models by targeting KEAP1

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

Ferroptosis, a form of cell death characterized by lipid peroxidation, is involved in neurodegenerative diseases such as Alzheimer´s disease (AD). Recent studies have shown that a first-line antimalarial drug artemisinin is effective to counteract AD pathology. In this study, we investigated the protective effect of artemisinin against neuronal ferroptosis and the underlying mechanisms. In hippocampal HT22 cells, pretreatment with artemisinin dose-dependently protected against Erastin-induced cell death with an EC50 value of 5.032 µM, comparable to the ferroptosis inhibitor ferrostatin-1 (EC50 = 4.39 µM). We demonstrated that artemisinin (10 μM) significantly increased the nuclear translocation of Nrf2 and upregulated SLC7A11 and GPX4 in HT22 cells. Knockdown of Nrf2, SLC7A11 or GPX4 prevented the protective action of artemisinin, indicating that its anti-ferroptosis effect is mediated by the Nrf2-SLC7A11-GPX4 pathway. Molecular docking and Co-Immunoprecipitation (Co-IP) analysis revealed that artemisinin competitively binds with KEAP1, promoting the dissociation of KEAP1-Nrf2 complex and inhibiting the ubiquitination of Nrf2. Intrahippocampal injection of imidazole-ketone-Erastin (IKE) induced ferroptosis in mice accompanied by cognitive deficits evidenced by lower preference for exploration of new objects and new object locations in the NOR and NOL tests. Artemisinin (5, 10 mg/kg, i.p.) dose-dependently inhibited IKE-induced ferroptosis in hippocampal CA1 region and ameliorated learning and memory impairments. Moreover, we demonstrated that artemisinin reversed Aβ1-42-induced ferroptosis, lipid peroxidation and glutathione depletion in HT22 cells, primary hippocampal neurons, and 3×Tg mice via the KEAP1-Nrf2 pathway. Our results demonstrate that artemisinin is a novel neuronal ferroptosis inhibitor that targets KEAP1 to activate the Nrf2-SLC7A11-GPX4 pathway.

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Fig. 1: Artemisinin opposes cysteine deprivation-induced ferroptosis.
Fig. 2: Artemisinin activates the Nrf2-SLC7A11-GPX4 signaling pathway.
Fig. 3: Artemisinin-mediated anti-ferroptosis activity relies on the Nrf2-SLC7A11-GPX4 signaling pathway.
Fig. 4: Artemisinin directly targets KEAP1 to activate Nrf2.
Fig. 5: Effect of artemisinin on ferroptosis in vivo.
Fig. 6: Artemisinin alleviated ferroptosis in Aβ1-42-treated neurons and 3×Tg mice.
Fig. 7: Schematic illustration of the main findings.

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Funding

This research was supported by National Natural Science Foundation of China (32070969 and 22274115), the Science and Technology Development Fund, Macao SAR (0104/2022/A2 and 0038/2020/AMJ), Guangdong, Hong Kong and Macao Joint Key Laboratory for New Drug Screening (EF2023-00054-FHS, GDSTC), the Guangdong Provincial Funding Committee for Basic and Applied Fundamental Research (2022-Natural Science Foundation, EF019/FHS-ZWH/2022, GDSTC), and University of Macau (MYRG-CRG2024-00019-FHS, MYRG-GRG2023-00118-FHS-UMDF and MYRG2022-00154-FHS).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Faculty of Health Sciences, University of Macau, Taipa, 999078, Macao, China

    Peng-xi Deng, Marta Silva, Xin Meng & Wen-hua Zheng

  2. Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health); Institute of Metabonomics & Medical NMR, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, China

    Peng-xi Deng & Hong-chang Gao

  3. Department of Cardiology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, 200040, China

    Na Yang

  4. Department of Neurology, Zhujiang Hospital of Southern Medical University, Guangzhou, 510280, China

    Qing Wang

  5. Faculty of Life and Health Sciences, Shenzhen Institutes of Advanced Technology (SIAT), Chinese Academy of Sciences, Shenzhen, 518055, China

    Xin Meng & Ke-qiang Ye

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  1. Peng-xi Deng
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Contributions

PXD performed the experiments and drafted the manuscript. NY, XM and QW performed part of experiments. WHZ and HCG conceived the hypothesis, designed the experiments. MS, KQY and WHZ revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hong-chang Gao or Wen-hua Zheng.

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Deng, Px., Silva, M., Yang, N. et al. Artemisinin inhibits neuronal ferroptosis in Alzheimer’s disease models by targeting KEAP1. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01378-6

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