Related Experiment Video
Updated: Oct 21, 2025
Evaluating Cell Death Signaling by Immunofluorescence in a Rat Model of Ischemic Stroke
Published on: January 3, 2025
Long non-coding RNA MEG3 regulates autophagy after cerebral ischemia/reperfusion injury
Tian-Hao Li1, Hong-Wei Sun1, Lai-Jun Song1
1Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China.
Abstract:
Severe cerebral ischemia/reperfusion injury has been shown to induce high-level autophagy and neuronal death. Therefore, it is extremely important to search for a target that inhibits autophagy activation. Long non-coding RNA MEG3 participates in autophagy. However, it remains unclear whether it can be targeted to regulate cerebral ischemia/reperfusion injury. Our results revealed that in oxygen and glucose deprivation/reoxygenation-treated HT22 cells, MEG3 expression was obviously upregulated, and autophagy was increased, while knockdown of MEG3 expression greatly reduced autophagy. Furthermore, MEG3 bound miR-181c-5p and inhibited its expression, while miR-181c-5p bound to autophagy-related gene ATG7 and inhibited its expression. Further experiments revealed that mir-181c-5p overexpression reversed the effect of MEG3 on autophagy and ATG7 expression in HT22 cells subjected to oxygen and glucose deprivation/reoxygenation. In vivo experiments revealed that MEG3 knockdown suppressed autophagy, infarct volume and behavioral deficits in cerebral ischemia/reperfusion mice. These findings suggest that MEG3 knockdown inhibited autophagy and alleviated cerebral ischemia/reperfusion injury through the miR-181c-5p/ATG7 signaling pathway. Therefore, MEG3 can be considered as an intervention target for the treatment of cerebral ischemia/reperfusion injury. This study was approved by the Animal Ethics Committee of the First Affiliated Hospital of Zhengzhou University, China (approval No. XF20190538) on January 4, 2019.
Insights
Long non-coding RNA MEG3 knockdown inhibits autophagy and reduces brain damage after cerebral ischemia/reperfusion injury by regulating the miR-181c-5p/ATG7 pathway, suggesting MEG3 as a therapeutic target.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Severe cerebral ischemia/reperfusion injury triggers significant autophagy and neuronal death.
- Identifying targets to inhibit autophagy is crucial for treating this condition.
- Long non-coding RNA MEG3 is implicated in autophagy, but its role in cerebral ischemia/reperfusion injury is unclear.
Purpose of the Study:
- To investigate the role of long non-coding RNA MEG3 in regulating autophagy in cerebral ischemia/reperfusion injury.
- To elucidate the underlying molecular mechanism involving microRNA-181c-5p and ATG7.
- To evaluate MEG3 as a potential therapeutic target for cerebral ischemia/reperfusion injury.
Main Methods:
- Utilized oxygen and glucose deprivation/reoxygenation models in HT22 cells and in vivo mouse models.
- Assessed MEG3 expression, autophagy levels, and neuronal death.
- Performed knockdown and overexpression experiments for MEG3 and miR-181c-5p.
- Investigated the binding interactions between MEG3, miR-181c-5p, and ATG7 using molecular biology techniques.
- Measured infarct volume and behavioral deficits in mice.
Main Results:
- MEG3 expression was upregulated, and autophagy was increased in HT22 cells under oxygen and glucose deprivation/reoxygenation.
- Knockdown of MEG3 significantly reduced autophagy and ATG7 expression.
- MEG3 directly bound and inhibited miR-181c-5p, which in turn bound and inhibited ATG7.
- Overexpression of miR-181c-5p reversed the effects of MEG3 on autophagy and ATG7.
- MEG3 knockdown in mice reduced autophagy, infarct volume, and improved behavioral deficits.
Conclusions:
- MEG3 knockdown alleviates cerebral ischemia/reperfusion injury by inhibiting autophagy via the miR-181c-5p/ATG7 pathway.
- MEG3 serves as a crucial regulator of autophagy in this injury model.
- MEG3 represents a promising therapeutic target for treating cerebral ischemia/reperfusion injury.
