Abstract
BackgroundAtherosclerosis is the main cause of carotid artery stenosis (CAS) which mostly occurs in the elderly. In this paper, the expression level of miR-375-3p in asymptomatic CAS patients and its diagnostic value for asymptomatic CAS were investigated, and the effects of miR-375-3p on the cell proliferation and migration of vascular smooth muscle cells (VSMCs) was further explored.Methods 98 healthy subjects and 101 asymptomatic CAS patients were participated in this study. qRT-PCR was used to measure the expression level of serum miR-375-3p, and the ROC curve was established to evaluate the predictive value of miR-375-3p for asymptomatic CAS. After transfection with miR-375-3p mimic or inhibitor in vitro, cell proliferation and migration were detected by CCK-8 assay, colony formation assay, and Transwell assay, respectively. The levels of TNF-α, IL-1β, IL-6 were detected by ELISA. Western blot was used to detect the protein expression of XIAP. Finally, luciferase reporter gene assay was applied to assess the interaction of miR-375-3p with target genes.ResultsThe expression level of serum miR-375-3p in asymptomatic CAS patients was significantly higher than that in healthy controls, and the AUC value of ROC curve was 0.888. The sensitivity and specificity were 80.2 and 86.7%, respectively, indicating that miR-375-3p had high diagnostic value for asymptomatic CAS. In vitro cell experiments showed that up-regulation of miR-375-3p significantly promoted the proliferation and migration of VSMCs, and also promoted the generation of inflammatory factors and phenotypic transformation of VSMCs. Luciferase reporter gene assay confirmed that XIAP was a target gene of miR-375-3p and was negatively regulated by miR-375-3p.ConclusionsIn this study, miR-375-3p may have a clinical diagnostic value for asymptomatic CAS patients which need further validation. Increased miR-375-3p levels in CAS may be associated with increased proliferation and migration of VSMCs via downregulation of the apoptosis inducing gene XIAP.
Highlights
Atherosclerosis is the main cause of carotid artery stenosis (CAS) which mostly occurs in the elderly
Inclusion criteria were based on previously published literature [20]: (1) the degree of ipsilateral internal carotid artery stenosis was greater than 50%, (2) asymptomatic status was determined by history review of patients, physical examination, and investigation of the National Institutes of Health Stroke Scale (NIH-SS)
Serum level of miR‐375‐3p Serum levels of miR-375-3p were significantly increased in asymptomatic CAS patients compared with healthy controls (Fig. 1, P < 0.001), indicating that the abnormal expression of miR-375-3p may be related to the occurrence of CAS
Summary
Atherosclerosis is the main cause of carotid artery stenosis (CAS) which mostly occurs in the elderly. The expression level of miR-375-3p in asymptomatic CAS patients and its diagnostic value for asymptomatic CAS were investigated, and the effects of miR-375-3p on the cell proliferation and migration of vascular smooth muscle cells (VSMCs) was further explored. Carotid artery stenosis (CAS) is one of the vital risk factors for CIEs, accounting for about 20% of all CIEs cases [2, 3]. CAS is considered to have no previous symptoms to be determined, and symptomatic CAS is determined to be associated with symptoms occurring in the preceding 6 months, while asymptomatic CAS could change from simple intimal thickening to symptomatic stenosis [4, 5]. Once asymptomatic CAS develops into symptomatic CAS, the incidence of cerebral infarction and myocardial infarction will be greatly increased. Early definite diagnosis of such lesions can provide reliable data for clinical indication of potential cerebral infarction patients and prevention of cerebral infarction
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