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Published on: May 6, 2014
Role of miR-15a-5p and miR-199a-3p in the inflammatory pathway regulated by NF-κB in experimental and human
Paula González-López1, Marta Álvarez-Villarreal1, Rubén Ruiz-Simón1
1Hepatic and Vascular Diseases Laboratory. Biochemistry and Molecular Biology Department, School of Pharmacy, Complutense University of Madrid, Madrid, Spain.
Background:
Cardiovascular diseases (CVDs) prevalence has significantly increased in the last decade and atherosclerosis development is the main trigger. MicroRNAs (miRNAs) are non-coding RNAs that negatively regulate gene expression of their target and their levels are frequently altered in CVDs.
Methods:
By RT-qPCR, we analysed miR-9-5p, miR-15a-5p, miR-16-5p and miR-199a-3p levels in aorta from apolipoprotein knockout (ApoE-/- ) mice, an experimental model of hyperlipidemia-induced atherosclerosis, and in human aortic and carotid atherosclerotic samples. By in silico studies, Western blot analysis and immunofluorescence studies, we detected the targets of the altered miRNAs.
Results:
Our results show that miR-15a-5p and miR-199a-3p are significantly decreased in carotid and aortic samples from patients and mice with atherosclerosis. In addition, we found an increased expression in targets of both miRNAs that participate in the inflammatory pathway of nuclear factor kappa B (NF-κB), such as IKKα, IKKβ and p65. In human vein endothelial cells (HUVECs) and vascular smooth muscle cells (VSMCs), the overexpression of miR-15a-5p or miR-199a-3p decreased IKKα, IKKβ and p65 protein levels as well as NF-κB activation. On the other hand, miR-15a-5p and miR-199a-3p overexpression reduced ox-LDL uptake and the inflammation regulated by NF-κB in VSMCs. Moreover, although miR-15a-5p and miR-199a-3p were significantly increased in exosomes from patients with advanced carotid atherosclerosis, only in the ROC analyses for miR-15a-5p, the area under the curve was 0.8951 with a p value of .0028.
Conclusions:
Our results suggest that the decrease of miR-199a-3p and miR-15a-5p in vascular samples from human and experimental atherosclerosis could be involved in the NF-κB activation pathway, as well as in ox-LDL uptake by VSMCs, contributing to inflammation and progression atherosclerosis. Finally, miR-15a-5p could be used as a novel diagnostic biomarker for advanced atherosclerosis.
Insights
Decreased miR-15a-5p and miR-199a-3p levels in atherosclerosis promote inflammation and vascular smooth muscle cell uptake of oxidized low-density lipoprotein. miR-15a-5p shows potential as a diagnostic biomarker for advanced atherosclerosis.
Area of Science:
- Cardiovascular Research
- Molecular Biology
- Biomarker Discovery
Background:
- Cardiovascular diseases (CVDs) are rising, with atherosclerosis as a primary driver.
- MicroRNAs (miRNAs) are key regulators of gene expression, often dysregulated in CVDs.
Purpose of the Study:
- Investigate the role of specific miRNAs (miR-9-5p, miR-15a-5p, miR-16-5p, miR-199a-3p) in atherosclerosis.
- Identify potential diagnostic biomarkers for atherosclerosis progression.
Main Methods:
- Analyzed miRNA levels in mouse and human atherosclerotic samples using RT-qPCR.
- Identified miRNA targets via in silico, Western blot, and immunofluorescence studies.
- Assessed miRNA effects on inflammatory pathways and oxidized LDL uptake in cell cultures.
Main Results:
- miR-15a-5p and miR-199a-3p were significantly decreased in atherosclerotic tissues.
- These miRNAs target components of the nuclear factor kappa B (NF-κB) inflammatory pathway.
- Overexpression of miR-15a-5p/miR-199a-3p reduced NF-κB activation and oxidized LDL uptake.
- miR-15a-5p in exosomes showed diagnostic potential for advanced atherosclerosis (AUC=0.8951).
Conclusions:
- Reduced miR-15a-5p and miR-199a-3p contribute to atherosclerosis via NF-κB activation and increased oxidized LDL uptake.
- miR-15a-5p is a promising diagnostic biomarker for advanced atherosclerosis.
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