Vitexin alleviates cerebral ischemia/reperfusion injury by regulating mitophagy via the SIRT1/PINK1/Parkin pathway

IF 3.7 3区医学 Q2 NEUROSCIENCES

Brain Research Bulletin Pub Date : 2025-05-27 DOI:10.1016/j.brainresbull.2025.111404

Chao Chen , Zhenzhong Zhang , Baolin Du , Chenling Lv

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引用次数: 0

Abstract

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.

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牡荆素通过SIRT1/PINK1/Parkin通路调节线粒体自噬，减轻脑缺血/再灌注损伤

目的探讨牡荆素通过调节线粒体自噬改善脑缺血再灌注损伤的保护作用及其机制。方法采用大脑中动脉闭塞再灌注法诱导小鼠局灶性CIRI。2,3,5-三苯四唑氯染色评价脑梗死。分别用神经功能缺损评分和苏木精-伊红染色评估神经功能缺损和脑组织损伤。HT22细胞进行氧-葡萄糖剥夺/再氧化（OGD/R）暴露以建立体外模型。在OGD/R之前，我们用vitexin、线粒体自噬抑制剂（Mdivi-1）或SIRT1抑制剂（EX-527）预处理HT22细胞。分别通过细胞计数试剂盒-8法和流式细胞术检测细胞活力和细胞凋亡。JC-1荧光染色和MitoSOX™Red染色分别用于评估线粒体膜电位（MMP）和检测线粒体活性氧（mtROS）水平。Western blot检测b细胞淋巴瘤2 （Bcl-2）、Bcl-2相关X蛋白（Bax）、微管相关蛋白1 A/ 1b轻链3 （LC3）、sequestosomes -1 （p62）、pten诱导激酶1 （PINK1）、Parkin以及沉默信息调节因子2 1 （SIRT1）的表达。结果我们的体内和体外实验证实，vitexin可显著缓解小鼠CIRI，减轻OGD/R暴露引起的HT22细胞损伤，并伴有线粒体自噬和SIRT1/PINK1/Parkin通路的激活。OGD/R+牡荆素+Mdivi-1组（与OGD/R+牡荆素组相比）显示细胞活力降低，凋亡增加，Bcl-2/Bax比值降低，MMP降低，mtROS水平升高，PINK1， LC3-II和Parkin表达下调，p62表达上调。同样，OGD/R+Vitexin+EX-527组细胞活力降低，凋亡增加，Bcl-2/Bax比值降低，MMP降低，mtROS水平升高，SIRT1、PINK1、LC3-II和Parkin表达下调，p62表达上调。结论牡荆素通过SIRT1/PINK1/Parkin通路激活线粒体自噬，改善CIRI。

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来源期刊

Brain Research Bulletin 医学-神经科学

CiteScore

6.90

自引率

2.60%

发文量

253

审稿时长

67 days

期刊介绍： The Brain Research Bulletin (BRB) aims to publish novel work that advances our knowledge of molecular and cellular mechanisms that underlie neural network properties associated with behavior, cognition and other brain functions during neurodevelopment and in the adult. Although clinical research is out of the Journal''s scope, the BRB also aims to publish translation research that provides insight into biological mechanisms and processes associated with neurodegeneration mechanisms, neurological diseases and neuropsychiatric disorders. The Journal is especially interested in research using novel methodologies, such as optogenetics, multielectrode array recordings and life imaging in wild-type and genetically-modified animal models, with the goal to advance our understanding of how neurons, glia and networks function in vivo.