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Published on: May 26, 2023
Vitexin alleviates cerebral ischemia/reperfusion injury by regulating mitophagy via the SIRT1/PINK1/Parkin pathway
Chao Chen1, Zhenzhong Zhang2, Baolin Du3
1Department of Cardiology, Hangzhou Hospital of Traditional Chinese Medicine, NO. 453 Stadium Road, Hangzhou, Zhejiang 310007, China.
Objective:
This study was conducted to elucidate vitexin's protective effects and underlying mechanism in ameliorating cerebral ischemia/reperfusion injury (CIRI) through regulation of mitophagy.
Methods:
Focal CIRI in mice was induced using the middle cerebral artery occlusion and reperfusion method. 2,3,5-triphenyltetrazolium chloride staining was performed for the evaluation of cerebral infarction. Neurological deficits and brain tissue damage were assessed by neurological deficit scores and hematoxylin-eosin staining, respectively. HT22 cells underwent oxygen-glucose deprivation/reoxygenation (OGD/R) exposure to develop an in vitro model. Prior to OGD/R, we pretreated the HT22 cells with vitexin, the mitophagy inhibitor (Mdivi-1), or the SIRT1 inhibitor (EX-527). Determination of cell viability and apoptosis were carried out through the cell counting kit-8 assay and flow cytometry, respectively. JC-1 fluorescence staining and MitoSOX™ Red staining were respectively performed for assessing mitochondrial membrane potential (MMP) and detecting levels of mitochondrial reactive oxygen species (mtROS). Expression of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), microtubule-associated protein 1 A/1B-light chain 3 (LC3), sequestosome-1 (p62), PTEN-induced kinase 1 (PINK1), Parkin, as well as silent information regulator two 1 (SIRT1) was determined via Western blot.
Results:
Vitexin was found to significantly alleviate CIRI in mice and mitigate HT22 cell injury due to OGD/R exposure, as confirmed by our in vivo and in vitro experiments, accompanied by activation of mitophagy and the SIRT1/PINK1/Parkin pathway. The OGD/R+Vitexin+Mdivi-1 group (versus the OGD/R+Vitexin group) displayed decreased cell viability, increased apoptosis, a reduced Bcl-2/Bax ratio, diminished MMP, elevated mtROS levels, downregulated PINK1, LC3-II, and Parkin expression, and upregulated p62 expression. Similarly, the OGD/R+Vitexin+EX-527 group showed reduced cell viability, increased apoptosis, a decreased Bcl-2/Bax ratio, decreased MMP, elevated mtROS levels, downregulated SIRT1, PINK1, LC3-II, and Parkin expression, and upregulated p62 expression.
Conclusion:
Vitexin ameliorates CIRI by activating mitophagy via the SIRT1/PINK1/Parkin pathway.
Insights
Vitexin protects against cerebral ischemia/reperfusion injury (CIRI) by activating mitophagy. This process involves the SIRT1/PINK1/Parkin pathway, crucial for cellular recovery and reducing brain damage.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Cerebral ischemia/reperfusion injury (CIRI) is a major cause of stroke-related brain damage.
- Mitophagy, the selective removal of damaged mitochondria, plays a critical role in cellular homeostasis and injury response.
- Vitexin, a natural flavonoid, has shown potential therapeutic benefits, but its mechanism in CIRI remains to be fully elucidated.
Purpose of the Study:
- To investigate the protective effects of vitexin against CIRI.
- To elucidate the underlying mechanism of vitexin's action, focusing on the regulation of mitophagy.
- To explore the role of the SIRT1/PINK1/Parkin pathway in vitexin-mediated neuroprotection.
Main Methods:
- In vivo study: Focal CIRI induced in mice via middle cerebral artery occlusion and reperfusion.
- In vitro study: HT22 cells subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to model CIRI.
- Assessment of cerebral infarction, neurological deficits, cell viability, apoptosis, mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (mtROS), and protein expression (Bcl-2, Bax, LC3, p62, PINK1, Parkin, SIRT1).
Main Results:
- Vitexin significantly alleviated CIRI in mice and protected HT22 cells from OGD/R-induced injury.
- Vitexin treatment activated mitophagy and the SIRT1/PINK1/Parkin pathway.
- Inhibition of mitophagy or SIRT1 abolished the protective effects of vitexin, leading to increased cell death and mitochondrial dysfunction.
Conclusions:
- Vitexin ameliorates CIRI by activating mitophagy through the SIRT1/PINK1/Parkin pathway.
- Targeting mitophagy represents a promising therapeutic strategy for managing CIRI.
- Vitexin holds potential as a therapeutic agent for stroke and other conditions involving CIRI.
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