Abstract
Oxidative stress and mitochondrial dysfunction are involved in cerebral ischemia/reperfusion injury-induced neuronal apoptosis. Mitophagy is the main method to eliminate dysfunctional mitochondria. Apelin-36, a type of neuropeptide, has been reported to exert protective effects in cerebral I/R (I/R) injury, but its precise mechanisms remain to be elucidated. To study the effects of Apelin-36 on oxidative stress and mitochondrial dysfunction in cerebral I/R injury, the oxygen-glucose deprivation/reperfusion (OGD/R) model with 6 h of ischemia and 6 h of reperfusion was established in HT22 cells. Results demonstrated that Apelin-36 protected against OGD/R injury by improving cell viability, decreasing the apoptotic cells ratio and increasing the ratio of Bcl-2/Bax. In addition, Apelin-36 treatment inhibited oxidative stress by downregulating the level of reactive oxygen species (ROS) and malondialdehyde (MDA) as well as the expression of inducible nitric oxide synthase (iNOS). And Apelin-36 also activated the level of superoxide dismutase (SOD) and glutathione (GSH). Mitochondrial apoptosis was also alleviated with Apelin-36 treatment detected by the mitochondrial membrane potential (MMP) and the expression of Cytochrome c (Cyt c), Cleaved caspase-9, and Cleaved caspase-3. Furthermore, the SIRT1-mediated PINK1/Parkin-dependent mitophagy was activated by Apelin-36 treatment with the downregulation of p62 and upregulation of LC3B-II and Beclin1. Both EX527 and Cyclosporine A (CsA), which are inhibitors of SIRT1 and mitophagy, markedly alleviated the inhibition of oxidative stress and mitochondrial dysfunction caused by Apelin-36. These findings suggest that SIRT1-mediated PINK1/Parkin-dependent mitophagy is involved in the neuroprotective effects of Apelin-36 on OGD/R-induced oxidative stress and mitochondrial dysfunction.
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The study was supported by the National Natural Science Foundation of China, Nos. 81870948 (to BB), 81671276 (to BHC), 81501018 (to CMW); the Natural Science Foundation of Shandong Province of China, No. ZR2014HL040 (to BHC); and Program Supporting Foundation for Teachers’ Research of **ing Medical University of China, No. JYFC2018KJ003 (to SSD). The funding sources had no role in study conception and design, data analysis or interpretation, paper writing, or deciding to submit this paper for publication.
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Study design: Baohua Cheng, Bo Bai; experimental implementation: Ziqi Shao, Shanshan Dou, Junge Zhu; data analysis: Ziqi Shao, Huiqing Wang, Dandan Xu, Chunmei Wang; manuscript writing: Ziqi Shao. All authors read and approved the final manuscript.
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Shao, Z., Dou, S., Zhu, J. et al. Apelin-36 Protects HT22 Cells Against Oxygen-Glucose Deprivation/Reperfusion-Induced Oxidative Stress and Mitochondrial Dysfunction by Promoting SIRT1-Mediated PINK1/Parkin-Dependent Mitophagy. Neurotox Res 39, 740–753 (2021). https://doi.org/10.1007/s12640-021-00338-w
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DOI: https://doi.org/10.1007/s12640-021-00338-w