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ATG5 Knockdown Attenuates Ischemia‒Reperfusion Injury by Reducing Excessive Autophagy-Induced Ferroptosis

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Abstract

Autophagy has been described to be both protective and pathogenic in cerebral ischemia/reperfusion (I/R) injury. The underlying association between autophagy and ferroptosis in ischemic stroke has not yet been clearly investigated. The purpose of this study was to explore the role of autophagy-related gene 5 (ATG5) in experimental ischemic stroke. After injection of ATG5 shRNA lentivirus, mice underwent surgery for transient middle cerebral artery occlusion (MCAO)-induced focal cerebral ischemia. The infarct volume, neurological function, apoptosis, reactive oxygen species (ROS), autophagy, and ferroptosis levels were evaluated. After MCAO, ATG5-knockdown mice had a smaller infarct size and fewer neurological deficits than wild-type mice. The levels of apoptosis and ROS in ischemic mouse brains were alleviated through ATG5 knockdown. The expression of LC3 I/II was reduced through ATG5 knockdown after MCAO. Additionally, the expression of beclin1 and LC3 II was increased after I/R, but the increase was counteracted by preconditioning with ATG5 knockdown. After ischemic stroke, the levels of Fe2+ and malondialdehyde (MDA) were increased, but they were reduced by ATG5 knockdown. Similarly, the expression of glutathione peroxidase 4 (GPX4) and glutathione (GSH) was decreased by I/R but elevated by ATG5 knockdown. The present study shows that ATG5 knockdown attenuates autophagy-induced ferroptosis, which may offer a novel potential approach for ischemic stroke treatment.

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The data presented in the study are included in the article/Supplementary Material, and further inquiries can be directed to the corresponding authors.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Nos. 81870939 and 82171336 to Xiaoxing Xiong; Nos. 82071339 and 82271370 to Lijuan Gu).

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  1. Hua Zhu and Yi Zhong contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, China

    Hua Zhu, Yi Zhong, Ran Chen, Lei Wang, Zhihong Jian & Xiaoxing Xiong

  2. Department of Neurosurgery, The Affiliated Huzhou Hospital, Zhejiang University School of Medicine (Huzhou Central Hospital), Huzhou, China

    Lei Wang, Yuntao Li & Xiaoxing Xiong

  3. Central Laboratory, Renmin Hospital of Wuhan University, Wuhan, China

    Lijuan Gu & Xiaoxing Xiong

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  1. Hua Zhu
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Contributions

Xiaoxing Xiong and Lijuan Gu designed this work. Hua Zhu and Yi Zhong wrote the manuscript. Hua Zhu, Ran Chen, Yuntao Li, Ran Chen, Lei Wang, Zhihong Jian, and Yi Zhong performed the experiments and analyzed the data. All authors contributed to manuscript revision and read and approved the submitted version.

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Correspondence to Lijuan Gu or Xiaoxing Xiong.

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Zhu, H., Zhong, Y., Chen, R. et al. ATG5 Knockdown Attenuates Ischemia‒Reperfusion Injury by Reducing Excessive Autophagy-Induced Ferroptosis. Transl. Stroke Res. 15, 153–164 (2024). https://doi.org/10.1007/s12975-022-01118-0

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