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Irisin Attenuates Apoptosis Following Ischemia–Reperfusion Injury Through Improved Mitochondria Dynamics and ROS Suppression Mediated Through the PI3K/Akt/mTOR Axis

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Abstract   

Irisin is a muscle-derived hormone that promotes the survival of motor neurons and enhances muscle size following injury. In this study, we investigated the beneficial effects and mechanism(s) of action of irisin in response to cerebral ischemia–reperfusion injury (CIRI). Right-middle cerebral artery occlusion (MCAO) and hypoxia/reoxygenation (H/R) models were generated in C57BL/6 J mice. Mouse neuronal cell lines (NSC-34) were used to confirm the molecular mechanisms of the protection afforded by irisin in response to CIRI. We found that irisin (250 μg/kg) improved cerebral function and reduced the cerebral infarct volume following CIRI. Irisin also protected neuronal cells against ischemia–reperfusion (I/R) induced apoptosis, assessed via TUNEL, and cleaved Caspase-3 staining. Western blotting of neuronal tissue from irisin treated I/R mice showed lower expression of pro-apoptotic Bax and caspase-9 (P < 0.001 and P < 0.01) and increased levels of the pro-survival protein Bcl-2 (P < 0.01 & P < 0.001 vs. I/R). Irisin also reduced the levels of reactive oxygen species (ROS) characterized through malondialdehyde (MDA) assays. Irisin was found to maintain mitochondrial homeostasis through the suppression of mitochondrial fission-linked dynamin-related protein 1 in CIRI mice (P < 0.01 and P < 0.05 v. I/R cohort). Moreover, mitochondrial fusion–related protein (Mfn2) and Opa1 expression were rescued following irisin treatment (P < 0.001 and P < 0.01 v. I/R cohort). Cell-based assays showed that irisin activates PI3K/AKT/mTOR signaling in the neurons of CIRI mice. Furthermore, the beneficial effects of irisin on NSC-34 cell-survival, mitochondrial function, and ROS generation were reversed by VS-5584, a highly specific PI3K/AKT/mTOR inhibitor. Collectively, these data highlight the ability of irisin to alleviate CIRI in vivo and in vitro. The mechanisms of action of irisin include the attenuation of apoptosis through the prevention of mitochondrial fission and increased mitochondrial fusion and the alleviation of oxidative stress through activation of the PI3K/AKT/mTOR axis. We therefore identify irisin as a much-needed therapeutic for CIRI.

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Data Availability

Data supporting the findings of this study are available upon reasonable request from the corresponding author.

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Funding

This study was supported by the National Natural Science Foundation of China (31870335); the Natural Science Foundation of Gansu Provincial Department of Science and Technology (20JR5RA344); the Gansu Province Health Industry Scientific Research Project (GSWSKY2021-017); the Lanzhou City Chengguan District Science and Technology Project (2021–9-3); and the “Cuiying Technology Innovation” Planning Project of Lanzhou University Second Hospital (CY2021-MS-B01).

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Authors and Affiliations

  1. Department of Neurology, Lanzhou University Second Hospital, No.82, Cuiyingmen, Lanzhou, 730030, Gansu, China

    Ji-fei Liu, Li-xia Chen, Juan-ping Zhou, Juan Gao, Jia-jia Zhang, Qiong-hui Wu, Wei Chen, De-yi Chen & Zhen-chang Zhang

  2. Institute of Genetics, School of Basic Medical Sciences, Lanzhou University, Lanzhou, 730000, Gansu, China

    Gang Su

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  1. Ji-fei Liu
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Contributions

JFL, GS, and ZCZ designed the experiments; LXC, JPZ, JG, JJZ, and QHW performed the experiments; WC, DYC, and ZCZ analyzed the data; JFL and GS prepared the manuscript. All authors read and approved submission of the manuscript.

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Correspondence to Zhen-chang Zhang.

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All in vivo protocols were accepted by the Institutional Animal Care and Use Committee of the Ethics Committee of Lanzhou University Second Hospital, according to regulations imposed by the China Council on Animal Care and Use.

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Liu, Jf., Su, G., Chen, Lx. et al. Irisin Attenuates Apoptosis Following Ischemia–Reperfusion Injury Through Improved Mitochondria Dynamics and ROS Suppression Mediated Through the PI3K/Akt/mTOR Axis. Mol Neurobiol 60, 4261–4272 (2023). https://doi.org/10.1007/s12035-023-03336-5

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