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Knockout of Rnf213 Ameliorates Cerebral Ischemic-reperfusion Injury by Inhibiting Neuronal Apoptosis Through the
Shumeng Li1, Yiheng Li2, Pengcheng Huang1
1Department of Neurology, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China.
Abstract:
Previous studies by us and others have shown that RING finger protein 213 (RNF213) is associated with cerebrovascular disease and systemic vasculopathy. Indeed, Rnf213 mRNA expression is increased in cerebral ischemia reperfusion injury (CIRI). The purpose of the present study was to investigate the role of Rnf213 in CIRI. Using the middle cerebral artery occlusion (MCAO) model, we confirmed that the expression of RNF213 protein was significantly upregulated in neurons in the ischemic penumbra. Rnf213 knockout mice were successfully generated using CRISPR/Cas9 technology. According to TTC staining and Bederson neurological scale, removal of Rnf213 decreased brain infarct volume and improved neurological deficit score, although the restoration of cerebral blood flow after MCAO was similar in WT and Rnf213-/- mice. In addition, the levels of p-Akt, p-GSK-3β, β-catenin and Bcl-2 were significantly increased 24 h after MCAO in the ischemic penumbra of the Rnf213-/- mice compared to WT mice, indicating that Rnf213 removal may ameliorate neuronal apoptosis by regulating the Akt/GSK-3β/β-catenin/Bcl-2 signaling pathway. Taken together, our study reveals that Rnf213 regulates neuronal apoptosis in CIRI, therefore impacting on brain infarct volume in brain ischemia.
Insights
Removing RING finger protein 213 (RNF213) reduces brain damage and improves outcomes in cerebral ischemia reperfusion injury. This suggests RNF213 plays a key role in neuronal apoptosis during stroke.
Area of Science:
- Neuroscience
- Molecular Biology
- Pathology
Background:
- RING finger protein 213 (RNF213) is linked to cerebrovascular and systemic vasculopathies.
- RNF213 expression increases in cerebral ischemia reperfusion injury (CIRI).
Purpose of the Study:
- To investigate the specific role of RNF213 in CIRI.
- To determine the impact of RNF213 on neuronal apoptosis and brain infarct volume.
Main Methods:
- Utilized the middle cerebral artery occlusion (MCAO) model in mice.
- Generated Rnf213 knockout mice using CRISPR/Cas9 technology.
- Assessed brain infarct volume (TTC staining) and neurological deficits (Bederson scale).
Main Results:
- RNF213 protein was upregulated in the ischemic penumbra.
- Rnf213 knockout mice exhibited reduced brain infarct volume and improved neurological scores post-MCAO.
- Knockout mice showed increased levels of p-Akt, p-GSK-3β, β-catenin, and Bcl-2 in the ischemic penumbra.
Conclusions:
- RNF213 removal ameliorates neuronal apoptosis in CIRI.
- The Akt/GSK-3β/β-catenin/Bcl-2 pathway is implicated in RNF213's role in CIRI.
- RNF213 is a regulatory factor in brain infarct volume during ischemia.
