MicroRNA-486-5p functions as a diagnostic marker for carotid artery stenosis and prevents endothelial dysfunction through inhibiting inflammation and oxidative stress

Abstract

Carotid artery stenosis (CAS) can cause ischemic stroke, and clinical intervention for CAS is critical clinically. The purpose of this study was to explore the expression changes of microRNA-486-5p in the serum of patients with CAS and its possible mechanism. Ninety-one cases with asymptomatic CAS were recruited, and serum levels of miR-486-5p were measured using RT-qPCR. The diagnostic ability was evaluated by drawing the receiver operating characteristic (ROC) curve. Human aortic endothelial cells (HAECs) were treated with oxidized low-density lipoprotein (oxLDL) to establish cell model, and cell proliferation and apoptosis were tested. The markers of cell inflammation and oxidative stress were detected via ELISA. The target gene was analyzed using bioinformatics analysis combined with luciferase reporting assay. CAS cases exhibited significantly low serum miR-486-5p levels in comparison with the control group and can identify asymptomatic CAS. Serum miR-486-5p manifested a negative correlation with the degree of carotid stenosis. Underexpression of miR-486-5p was also detected in ox-LDL treated HAECs. OxLDL treatment contributes to inflammatory response and oxidative stress of HAECs; however, these adverse impacts caused by ox-LDL were reversed by miR-486-5p upregulation. NFAT5 was confirmed to be the target gene of miR-486-5p in HAECs. MiR-486-5p serves as a promising biomarker for the early identification of CAS. Overexpression of miR-486-5p can prevent endothelial dysfunction, and the mechanism might be related to anti-inflammation and anti-oxidation via targeting NFAT5.

Keywords: Carotid artery stenosis; endothelial dysfunction; inflammation; miR-486-5p; oxidative stress.

Graphical abstract

Figure 1.

The diagnostic value of dysregulation of miR-486-5p in CAS patients. A. CAS cases exhibited significantly low serum miR-486-5p levels when compared with the control group (P < 0.001). B. ROC curve of serum miR-486-5p for identifying CAS from healthy individuals.

Figure 2.

In ox-LDL treated cells, low levels of miR-486-5p were also detected, which was reversed by miR-486-5p mimic transfection (a). MiR-486-5p negatively regulated the HAECs apoptosis (b), inflammation (c) and oxidative stress (d) induced by ox-LDL. *** P < 0.001 (VS control group); ^### P < 0.001 (VS ox-LDL group).

Figure 3.

Target relationship confirmation between miR-486-5p and NFAT5. A. TargetScan showed a target binding sequence between miR-486-5p and NFAT5. B. The luciferase activity of HAECs was regulated by miR-486-5p mimic or inhibitor. C. The mRNA levels of NFAT5 in ox-LDL treated cells transfected with miR- miR-486-5p mimic or inhibitor. *** P < 0.001 (VS control group); ^### P < 0.001 (VS ox-LDL group).