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miR-495-3p attenuates cerebral ischemia-reperfusion-induced neuronal inflammation and apoptosis by targeting CCL2
XiaoDong Yu1, LiZhi Xue1, WenQin Zou1
1Department of Neurology, Shiyan Renmin Hospital, No. 39 Chaoyang Middle Road, Maojian District, Shiyan City, 442000, Hubei Province, China.
Abstract:
This study aimed to explore the function of miR-495-3p in cerebral ischemia-reperfusion injury (CI/RI) and reveal its potential molecular mechanism. In vivo and in vitro models of CI/RI were established by MACO/R and OGD/R, respectively. Neural function scores, HE staining, and TUNEL staining assessed the degree of brain tissue injury in mice. LDH assay, MTT assay, and flow cytometry evaluated neuronal toxicity, viability, and apoptosis rate. ELISA and Western blot evaluated inflammatory factors and the NF-κB pathway. Dual-luciferase reporting assay and RIP explored the targeting relationship between miR-495-3p and CCL2. miR-495-3p was abnormally low in MACO/R mouse brain tissue and OGD/R-damaged neurons, while CCL2 was highly expressed. miR-495-3p overexpression improved neuronal apoptosis and inflammation in the brain tissue of MACO/R mice. Consistent results were also obtained in in vitro experiments. Enhancing CCL2 or knocking down miR-495-3p aggravated OGD/R-induced neuronal damage. The deleterious effects of miR-495-3p knockdown were prevented by the knockdown of CCL2. miR-495-3p targeted CCL2. miR-495-3p improves CI/R-mediated neuronal apoptosis and inflammation through targeted regulation of CCL2 expression. These results provide data support for CI/RI-targeting drugs and the understanding of disease mechanisms.
Insights
MicroRNA-495-3p (miR-495-3p) protects against cerebral ischemia-reperfusion injury (CI/RI) by downregulating CCL2. This finding offers new therapeutic targets for stroke and related brain injuries.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cerebral ischemia-reperfusion injury (CI/RI) is a significant cause of neurological damage.
- The precise molecular mechanisms underlying CI/RI remain incompletely understood.
- MicroRNAs (miRNAs) have emerged as critical regulators in various pathological processes, including brain injury.
Purpose of the Study:
- To investigate the role of miR-495-3p in CI/RI.
- To elucidate the molecular mechanism by which miR-495-3p influences CI/RI.
- To identify potential therapeutic targets for CI/RI.
Main Methods:
- Establishment of in vivo (MACO/R) and in vitro (OGD/R) models of CI/RI.
- Assessment of brain injury using neural function scores, HE staining, and TUNEL staining.
- Evaluation of neuronal damage via LDH assay, MTT assay, and flow cytometry.
- Analysis of inflammatory factors and the NF-κB pathway using ELISA and Western blot.
- Determination of the miR-495-3p and CCL2 interaction using dual-luciferase reporting assay and RIP.
Main Results:
- miR-495-3p expression was significantly downregulated, while CCL2 expression was upregulated in CI/RI models.
- Overexpression of miR-495-3p attenuated neuronal apoptosis and inflammation in CI/RI models.
- CCL2 knockdown reversed the detrimental effects of miR-495-3p knockdown.
- miR-495-3p directly targets and inhibits CCL2 expression.
Conclusions:
- miR-495-3p plays a protective role in CI/RI by suppressing neuronal apoptosis and inflammation.
- The therapeutic effect of miR-495-3p is mediated through the targeted downregulation of CCL2.
- miR-495-3p represents a promising therapeutic target for managing CI/RI and related neurological disorders.
