Abstract
Cannabidiol (CBD) reportedly exerts protective effects against many psychiatric disorders and neurodegenerative diseases, but the mechanisms are poorly understood. In this study, we explored the molecular mechanism of CBD against cerebral ischemia. HT-22 cells or primary cortical neurons were subjected to oxygen-glucose deprivation insult followed by reoxygenation (OGD/R). In both HT-22 cells and primary cortical neurons, CBD pretreatment (0.1, 0.3, 1 μM) dose-dependently attenuated OGD/R-induced cell death and mitochondrial dysfunction, ameliorated OGD/R-induced endoplasmic reticulum (ER) stress, and increased the mitofusin-2 (MFN2) protein level in HT-22 cells and primary cortical neurons. Knockdown of MFN2 abolished the protective effects of CBD. CBD pretreatment also suppressed OGD/R-induced binding of Parkin to MFN2 and subsequent ubiquitination of MFN2. Overexpression of Parkin blocked the effects of CBD in reducing MFN2 ubiquitination and reduced cell viability, whereas overexpressing MFN2 abolished Parkin’s detrimental effects. In vivo experiments were conducted on male rats subjected to middle cerebral artery occlusion (MCAO) insult, and administration of CBD (2.5, 5 mg · kg−1, i.p.) dose-dependently reduced the infarct volume and ER stress in the brains. Moreover, the level of MFN2 within the ischemic penumbra of rats was increased by CBD treatment, while the binding of Parkin to MFN2 and the ubiquitination of MFN2 was decreased. Finally, short hairpin RNA against MFN2 reversed CBD’s protective effects. Together, these results demonstrate that CBD protects brain neurons against cerebral ischemia by reducing MFN2 degradation via disrupting Parkin’s binding to MFN2, indicating that MFN2 is a potential target for the treatment of cerebral ischemia.
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Acknowledgements
This work was supported by National Natural Science Foundation of China (Nos. 82173802 and 82273902), Science and Technology Program of Guangzhou (No. 202002030494), Guangdong Basic and Applied Basic Research Foundation (No. 2022A1515012317).
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BTX performed CCK-8 assay, immunoblotting, immunocytochemistry, co-IP, PCR, transfection, and partial animal experiments. MFL performed immunoblotting, PCR, and helped with animal experiments. KCC performed immunocytochemistry, PI staining and helped with animal experiments. XL performed immunoblotting and helped with cell culture. NBC performed animal experiments. BTX, MFL, XL, and KCC analyzed the data. HTW and JPX designed the experiments and supervised the project. BTX and HTW wrote the manuscript. All authors commented to the manuscript.
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Xu, Bt., Li, Mf., Chen, Kc. et al. Mitofusin-2 mediates cannabidiol-induced neuroprotection against cerebral ischemia in rats. Acta Pharmacol Sin 44, 499–512 (2023). https://doi.org/10.1038/s41401-022-01004-3
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DOI: https://doi.org/10.1038/s41401-022-01004-3
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