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FTO inhibits oxidative stress by mediating m6A demethylation of Nrf2 to alleviate cerebral ischemia/reperfusion injury

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Abstract

Current therapies are of limited efficacy in cerebral ischemia/reperfusion (I/R) injury. Based on the important role of oxidative stress in cerebral I/R injury, this study aimed to explore how the N6-adenosine methylation (m6A) demethylase FTO affects oxidative stress. Middle cerebral artery occlusion/reperfusion (MCAO/R)-induced rat model and oxygen and glucose deprivation/re-oxygenation (OGD/R)-induced SH-SY5Y cells were established as in vivo and in vitro model, respectively. The neurological score of rats was measured, and the volume of cerebral infarction was measured by TTC staining. The levels of FTO, nuclear factor-erythroid 2-related factor (Nrf2), and the activity of m6A demethylase FTO were detected. The m6A methylation level of Nrf2 mRNA was detected by MeRIP experiment. Flow cytometry and MTT assay were used to detect apoptosis and proliferation in vitro. TUNEL assay was used to detect apoptosis in brain tissues. FTO and Nrf2 expressions were decreased in the MCAO/R rat brain tissues and OGD/R SH-SY5Y cells, while the m6A methylation level of Nrf2 mRNA was significantly increased. Overexpression of FTO upregulated Nrf2 expression by decreasing the m6A methylation level of Nrf2 mRNA. m6A binding protein YT521-B homology (YTH) domain family protein 2 (YTHDF2) promoted the degradation of Nrf2 by promoting the m6A methylation level of Nrf2 mRNA. Furthermore, SH-SY5Y cell apoptosis was increased and cell viability was decreased after the addition of methyltransferases METTL 3/14, thus blocking FTO to protect SH-SY5Y cells from oxidative stress injury. In vivo, overexpression of FTO decreased the area of cerebral ischemia infarction and the extent of cell apoptosis. In conclusion, FTO increases Nrf2 expression by mediating m6A demethylation of Nrf2 mRNA, thereby inhibiting oxidative stress response and ultimately alleviating cerebral I/R injury.

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Funding

This study was supported by the Shandong Provincial Natural Science Foundation (ZR2013HQ016 and ZR2021MH059), the Key Research and Development Project of Shandong Province (2015GGH318011).

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Authors and Affiliations

  1. Department of Neurology, the First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, 16766 Jingshi Road, Jinan, 250014, People’s Republic of China

    Shuang Li, Shasha Li, Ru Wang, Mengke Zhao & Xiaomin Liu

  2. Institute of Traditional Chinese Medicine Pharmacology, Shandong Academy of Chinese Medicine, Jinan, 250014, People’s Republic of China

    Lijing Hou

Authors
  1. Lijing Hou
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  2. Shuang Li
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Contributions

Lijing Hou and Xiaomin Liu: conception, design, and analysis of data, performed the data analysis, and wrote the manuscript. Shuang Li, Shasha Li, Mengke Zhao, and Ru Wang: contributed to the conception of the study. Lijing Hou and Xiaomin Liu: contributed significantly to analysis and manuscript preparation. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Xiaomin Liu.

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Ethics approval

All animal experiments were approved by the Animal Ethics Committee of the First Affiliated Hospital of Shandong First Medical University (NO. 2021-S046).

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Key Points

1. The influence of FTO on cerebral infarction I/R injury revealed for the first time.

2. FTO mediated oxidative stress response by m6A demethylation of Nrf2 mRNA.

3. YTHDF2 binds to m6A methylated Nrf2 mRNA and participates in its degradation.

Supplementary Information

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ESM 1

Binding sites between YTHDF2 and Nrf2 mRNA predicted by RNAInter. (PNG 1593 kb)

High Resolution (TIF 537 kb)

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Hou, L., Li, S., Li, S. et al. FTO inhibits oxidative stress by mediating m6A demethylation of Nrf2 to alleviate cerebral ischemia/reperfusion injury. J Physiol Biochem 79, 133–146 (2023). https://doi.org/10.1007/s13105-022-00929-x

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