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miR‑183‑5p attenuates cerebral ischemia injury by negatively regulating PTEN
Li Zhu1, Xueying Zhou2, Shanshan Li3
1Department of Neurology, Qilu Hospital of Shandong University and Brain Science Research Institute, Shandong University, Jinan, Shandong 250012, P.R. China.
Abstract:
Cerebral ischemia is a common cerebrovascular disease caused by the occlusion of a cerebral blood vessel. MicroRNAs (miRNAs/miRs) are emerging regulators of various human diseases, including cerebral ischemia. Upregulation of miR‑183‑5p has been reported to alleviate liver injury induced by ischemia‑reperfusion (I/R). However, the effect of miR‑183‑5p on cerebral ischemia injury remains unknown. The present study evaluated the effects of miR‑183‑5p on ischemia injury using ischemic models of mouse brains exposed to transient middle cerebral artery occlusion and Neuro‑2A (N2A) neuroblastoma cells exposed to oxygen‑glucose‑deprivation (OGD) and subsequently reoxygenated. Ischemia was evaluated in mice using neurological function scores, cerebral edema, 2,3,5‑triphenyltetrazoliumchloride, Nissl and Fluoro‑Jade B staining assays. In addition, miR‑183‑5p expression, N2A cell viability and the expression levels of apoptosis‑associated proteins were detected by quantitative PCR, Cell Counting Kit‑8 assay, flow cytometry and western blotting. The association between miR‑183‑5p and phosphatase and tensin homolog (PTEN) was also confirmed by a luciferase reporter assay. The results revealed that miR‑183‑5p expression was decreased and brain damage was increased in ischemic mice compared with the sham group. Additionally, miR‑183‑5p levels were reduced, and apoptosis was increased in N2A cells exposed to ischemia compared with the control group. Following transfection with agomiR‑183‑5p, cerebral ischemic injury and apoptosis levels were reduced in the in vivo I/R stroke model and OGD‑induced N2A cells. In addition, PTEN was determined to be a target of miR‑183‑5p following elucidation of a direct binding site. Overexpression of PTEN reversed the miR‑183‑5p‑induced N2A cell apoptosis inhibition and survival after OGD. The results of the present study suggested that miR‑183‑5p reduced ischemic injury by negatively regulating PTEN, which may aid the development of a novel therapeutic strategy for cerebral ischemia.
Insights
MicroRNA-183-5p (miR-183-5p) protects against cerebral ischemia injury by downregulating PTEN. This finding offers a potential new therapeutic strategy for stroke and related conditions.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cerebral ischemia, a cerebrovascular disease from blood vessel occlusion, poses significant health risks.
- MicroRNAs (miRNAs) are key regulators in various diseases, including cerebral ischemia, but the role of miR-183-5p is unclear.
- Previous studies indicated miR-183-5p alleviates liver ischemia-reperfusion injury, suggesting potential neuroprotective roles.
Purpose of the Study:
- To investigate the effect of miR-183-5p on cerebral ischemia injury.
- To explore the underlying molecular mechanisms, including the potential target gene PTEN.
- To evaluate miR-183-5p as a therapeutic target for cerebral ischemia.
Main Methods:
- Established transient middle cerebral artery occlusion (MCAO) in mice and oxygen-glucose deprivation (OGD) in Neuro-2A (N2A) cells.
- Assessed neurological function, cerebral edema, cell viability, and apoptosis using various assays (e.g., TTC, Nissl, flow cytometry, Western blotting).
- Utilized quantitative PCR, luciferase reporter assays, and transfection with agomiR-183-5p to confirm miR-183-5p targets and effects.
Main Results:
- miR-183-5p expression was significantly decreased in ischemic mouse brains and OGD-treated N2A cells.
- Overexpression of miR-183-5p (using agomiR-183-5p) reduced brain damage, neurological deficits, and apoptosis in both in vivo and in vitro models.
- Phosphatase and tensin homolog (PTEN) was identified as a direct target of miR-183-5p, and PTEN overexpression counteracted the protective effects of miR-183-5p.
Conclusions:
- miR-183-5p exerts a protective effect against cerebral ischemia injury.
- This protective mechanism involves the negative regulation of PTEN by miR-183-5p.
- miR-183-5p represents a promising therapeutic target for developing novel treatments for cerebral ischemia.
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