Deciphering the neuroprotective mechanisms of RACK1 in cerebral ischemia‐reperfusion injury: Pioneering insights into mitochondrial autophagy and the PINK1/Parkin axis

• Published in: CNS neuroscience & therapeutics Vol. 30; no. 8; pp. e14836 - n/a
• Main Authors: Zhao, Lanqing, Chen, Yu, Li, Hongxi, Ding, Xiaoxu, Li, Jinwei
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01-08-2024; John Wiley and Sons Inc
• Subjects: Animals; Apoptosis; Apoptosis - physiology; Autophagy; Autophagy - physiology; Brain Ischemia - metabolism; Brain Ischemia - pathology; cell apoptosis; Cell Line, Tumor; Cell viability; Cerebral blood flow; Cerebral infarction; cerebral ischemia‐reperfusion injury; Cerebrovascular diseases; Cloning; CRISPR; Datasets; Gene expression; Genome editing; Glucose; Humans; Infarction, Middle Cerebral Artery - metabolism; Infarction, Middle Cerebral Artery - pathology; Ischemia; Kinases; Laboratory animals; Male; Mitochondria; Mitochondria - metabolism; mitochondrial autophagy; mitochondrial function; Molecular modelling; Neoplasm Proteins; Neurological diseases; Neuroprotection; Neuroprotection - physiology; Original; Parkin protein; PINK/Parkin pathway; Protein Kinases - genetics; Protein Kinases - metabolism; PTEN-induced putative kinase; RACK1; Rats; Rats, Sprague-Dawley; Receptors for Activated C Kinase - metabolism; Reperfusion; Reperfusion Injury - metabolism; Reperfusion Injury - pathology; Signal Transduction - physiology; Software; Transcriptomes; Ubiquitin-Protein Ligases - genetics; Ubiquitin-Protein Ligases - metabolism; Vascular diseases; Veins & arteries; China; mitochondrial function; cerebral ischemia‐reperfusion injury; neuroprotection; PINK/Parkin pathway; RACK1; cell apoptosis; mitochondrial autophagy
• ISSN: 1755-5930; 1755-5949; 1755-5949
• DOI: 10.1111/cns.14836

Introduction
Cerebral ischemia‐reperfusion injury (CIRI) is a common and debilitating complication of cerebrovascular diseases such as stroke, characterized by mitochondrial dysfunction and cell apoptosis. Unraveling the molecular mechanisms behind these processes is essential for developing effective CIRI treatments. This study investigates the role of RACK1 (receptor for activated C kinase 1) in CIRI and its impact on mitochondrial autophagy.
Methods
We utilized high‐throughput transcriptome sequencing and weighted gene co‐expression network analysis (WGCNA) to identify core genes associated with CIRI. In vitro experiments used human neuroblastoma SK‐N‐SH cells subjected to oxygen and glucose deprivation (OGD) to simulate ischemia, followed by reperfusion (OGD/R). RACK1 knockout cells were created using CRISPR/Cas9 technology, and cell viability, apoptosis, and mitochondrial function were assessed. In vivo experiments involved middle cerebral artery occlusion/reperfusion (MCAO/R) surgery in rats, evaluating neurological function and cell apoptosis.
Results
Our findings revealed that RACK1 expression increases during CIRI and is protective by regulating mitochondrial autophagy through the PINK1/Parkin pathway. In vitro, RACK1 knockout exacerbated cell apoptosis, while overexpression of RACK1 reversed this process, enhancing mitochondrial function. In vivo, RACK1 overexpression reduced cerebral infarct volume and improved neurological deficits. The regulatory role of RACK1 depended on the PINK1/Parkin pathway, with RACK1 knockout inhibiting PINK1 and Parkin expression, while RACK1 overexpression restored them.
Conclusion
This study demonstrates that RACK1 safeguards against neural damage in CIRI by promoting mitochondrial autophagy through the PINK1/Parkin pathway. These findings offer crucial insights into the regulation of mitochondrial autophagy and cell apoptosis by RACK1, providing a promising foundation for future CIRI treatments.
This study reveals the key role of RACK1 in CIRI. The study finds that RACK1 regulates mitochondrial function and autophagy. The study confirms the centrality of the PINK1/Parkin pathway. The study offers new strategies for treating CIRI. The study provides comprehensive in vitro and in vivo evidence.