摘要： 目的 通过检测颈动脉粥样硬化性狭窄患者外周血miRNA-210-5P相对表达量，分析 miRNA-210-5P及其靶基因功能。方法 选择2015年7月至2018年9月诊断明确的颈动脉粥样硬化性狭窄患者（CAS组，146例）作为观察对象，采用逆转录聚合酶链反应（RT-PCR）检测不同处理组受试者 外周血miRNA-210-5P相对表达量，并以TargetScan和CoMeTa数据库进行靶基因预测、DAVID数据库进行靶基因功能富集分析（GO分析）和KEGG信号通路分析。结果 与无颈动脉粥样硬化性狭窄患者或正常受试者（对照组，60例）相比，CAS组患者血清miRNA-210-5P相对表达量升高（t=14.759， P= 0.000）。其中，重度狭窄组（31例； q=23.028， P=0.000）、中度狭窄组（53例； q=6.657， P=0.000）、轻度狭窄组（62例； q=42.612， P=0.000）患者外周血miRNA-210-5P相对表达量均高于对照组；而中度狭窄组 （q=34.538， P=0.000）和重度狭窄组（q=11.914， P=0.000）miRNA-210-5P相对表达量高于轻度狭窄组； 重度狭窄组亦高于中度狭窄组（q=16.983， P=0.000）。ROC曲线显示，miRNA-210-5P预测颈动脉粥样硬化中至重度狭窄的曲线下面积为0.943，当最佳临界值为1.495时，其预测灵敏度90.33%、特异度 92.54%；生物信息学分析提示，miRNA-210-5P潜在靶基因包括 VEGFA、KCMF1、HMGCS1、KLF12、 EFNA3、GIT2等54个基因；GO分析显示，miRNA-210-5P靶基因功能主要富集于血管生成、神经元发育、 DNA转录因子活性的正性调控、内皮细胞趋化性、细胞迁移与分化黏附等；对KEGG信号通路的检测显 示，miRNA-210-5P靶基因主要富集于突触导向信号转导通路。结论 颈动脉粥样硬化性狭窄患者血清 miRNA-210-5P表达上调，且可能通过调控多种靶基因而作用于突触导向信号转导通路中，参与颈动脉粥样硬化性狭窄的发生与发展。

关键词: 颈动脉狭窄, 动脉粥样硬化, 微RNAs, 基因, 计算生物学

Abstract: Objective To investigate the relative expression of miRNA-210-5P in serum of patients with carotid atherosclerotic stenosis (CAS) and to explore the function of miRNA-210-5P and its target genes using bioinformatics methods. Methods We selected 146 patients with CAS from July 2015 to September 2018 in our hospital. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the relative expression of miRNA-210-5P in peripheral blood of all enrolled patients. The target genes were predicted by using TargetScan and CoMeTa databases. The target genes of miRNA-210-5P were enriched by Gene Ontology (GO) using DAVID data and were performed with KEGG Pathway analysis. Results Compared with non-CAS patients or normal subjects (control group, N = 60), the relative expression of miRNA-210-5P in the serum of CAS group was significantly increased(t=14.759, P=0.000). The relative expressions of serum miRNA-210-5P in severe stenosis group(N=31; q=23.028, P=0.000), moderate stenosis group (N=53; q=6.657, P=0.000) and mild stenosis group (N=62; q=42.612, P= 0.000) were higher than that in control group. The relative expressions of serum miRNA-210-5P in moderate stenosis group (q =34.538, P =0.000) and severe stenosis group (q =11.914, P =0.000) were significantly higher than that in mild stenosis group. There lative expressions of serum miRNA-210-5P in severe stenosis group was significantly higher than thatin moderate stenosis group (q=16.983, P=0.000). Receive roperating characteristic (ROC) curve showed that the miRNA-210-5P predicted the area under the curve (AUC) of moderate to severe stenosis in CAS to be 0.943, the sensitivity was 90.33% and the specificity was 92.54% at the best cutoff value of 1.495. Bioinformatics analysis showed there were 54 potential target genes of miRNA-210-5P, such as VEGFA, KCMF1, HMGCS1, KLF12, EFNA3, GIT2, etc. GO analysis showed that the target genes of miRNA-210-5P were involved in angiogenesis, neuronal development, positive regulation of DNA transcription factor activity, endothelial cell chemotaxis, cell migration and differentiation and adhesion processes. KEGG Pathway analysis displayed miRNA-210-5P target genes were mainly enriched in synaptic-directed factor signal transduction pathways. Conclusions The expression of miRNA-210-5P inperipheral blood of CAS patients is upregulated,and it may participate in the process of CAS occurrence and development by regulating multiple target genes and acting on synaptic-directed signaling pathways.

Key words: Carotid stenosis, Atherosclerosis, MicroRNAs, Genes, Computational biology