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miR-214-3p Promotes ox-LDL-Induced Macrophages Ferroptosis and Inflammation via GPX4
Xueliang Pei1, Facai Cui2, Yu Chen3
1Department of Cardiovascular Surgery, Fuwai Central China Cardiovascular Hospital, Zhengzhou, Henan, People's Republic of China.
Purpose:
Atherosclerosis (AS) is a chronic inflammatory disease caused by the dysregulation of lipid metabolism. It has been established that oxidized low-density lipoprotein (ox-LDL)-induced macrophage inflammation and ferroptosis play important roles in AS. However, the mechanism by which ox-LDL induces inflammation in macrophages requires further investigation.
Materials And Methods:
A foam cell model derived from ox-LDL-induced macrophages was constructed to study its mechanism of action. The levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α were evaluated using an Enzyme-Linked Immunosorbent Assay (ELISA). Oil Red O staining was used to detect intracellular lipid accumulation levels. Lactate dehydrogenase (LDH), malondialdehyde (MDA), reactive oxygen species (ROS), and Fe2+ levels were assessed. Dual-luciferase and RNA-binding protein immunoprecipitation (RIP) experiments validated the binding relationship between microRNA (miR)-214-3p and glutathione peroxidase 4 (GPX4).
Results:
The levels of IL-6, IL-1β, and TNF-α were significantly increased in ox-LDL-induced macrophages, accompanied by increased lipid accumulation, indicating the promotion of foam cell formation. Additionally, ox-LDL increased LDH, MDA, ROS, and Fe2+. The expression of miR-214-3p positively correlated with ox-LDL concentration in macrophages. Treatment with an miR-214-3p inhibitor reduces lipid accumulation, inflammatory responses, and ferroptosis in macrophages. Dual-luciferase and RIP experiments confirmed that miR-214-3p regulates GPX4 transcription. Silenced GPX4 reversed the inflammatory effects of the miR-214-3p inhibitor on ox-LDL-induced macrophages. Low GPX4 expression also increased lipid accumulation and ferroptosis in macrophages.
Conclusion:
miR-214-3p promotes macrophage ferroptosis and inflammation induced by ox-LDL. This mechanism may be associated with miR-214-3p-induced GPX4 expression, which underscores the therapeutic significance of targeting macrophage inflammation and ferroptosis in addressing AS.
Insights
MicroRNA-214-3p promotes macrophage inflammation and ferroptosis in atherosclerosis by regulating glutathione peroxidase 4. Targeting this pathway offers therapeutic potential for atherosclerosis.
Area of Science:
- Biomedical Science
- Molecular Biology
- Cardiovascular Research
Background:
- Atherosclerosis (AS) is a chronic inflammatory disease driven by lipid metabolism dysregulation.
- Oxidized low-density lipoprotein (ox-LDL)-induced macrophage inflammation and ferroptosis are key contributors to AS pathogenesis.
- The precise mechanisms by which ox-LDL triggers macrophage inflammation require elucidation.
Purpose of the Study:
- To investigate the role of microRNA-214-3p (miR-214-3p) in ox-LDL-induced macrophage inflammation and ferroptosis.
- To elucidate the molecular mechanism linking miR-214-3p, glutathione peroxidase 4 (GPX4), and AS development.
- To assess the therapeutic potential of targeting the miR-214-3p/GPX4 axis in AS.
Main Methods:
- Established a foam cell model using ox-LDL-induced macrophages.
- Quantified inflammatory markers (IL-6, IL-1β, TNF-α) via ELISA.
- Assessed intracellular lipid accumulation using Oil Red O staining.
- Measured markers of oxidative stress and ferroptosis (LDH, MDA, ROS, Fe2+).
- Validated the interaction between miR-214-3p and GPX4 using dual-luciferase and RIP assays.
Main Results:
- Ox-LDL significantly increased macrophage inflammation, lipid accumulation, and ferroptosis markers.
- miR-214-3p expression was upregulated by ox-LDL in macrophages.
- Inhibition of miR-214-3p attenuated inflammation, lipid accumulation, and ferroptosis.
- miR-214-3p directly targets and regulates GPX4 transcription.
- GPX4 knockdown reversed the protective effects of miR-214-3p inhibition, confirming its role.
Conclusions:
- miR-214-3p exacerbates ox-LDL-induced macrophage inflammation and ferroptosis.
- The miR-214-3p/GPX4 regulatory axis is a critical mechanism in AS pathogenesis.
- Targeting macrophage inflammation and ferroptosis via the miR-214-3p/GPX4 pathway holds therapeutic promise for atherosclerosis.
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