Wang Kai, Huang Xi-Tong, Miao Yan-Ping, Bai Xiao-Long, Jin Feng
Department of Neurosurgery, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.
Department of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, China.
Ann Transl Med. 2022 Nov;10(22):1201. doi: 10.21037/atm-22-3768.
Atherosclerosis (AS) seriously affects human health. The role of microRNAs (miRNAs) in the pathogenesis and progression of AS has become a focus of research. Our goal was to identify the biological effect of differentially expressed miRNAs (DE-miRNAs) in AS.
To analyze differentially expressed genes (DEGs), including differentially expressed mRNAs (DE-mRNAs) and DE-miRNAs, in AS by using the Gene Expression Omnibus (GEO) database and limma package. DEGs protein-protein interaction (PPI) network and functional enrichment analysis were constructed by using the search tool for the retrieval of interacting genes/proteins (STRING) database, Cytoscape software and Cytoscape plugin "ClueGO2.5.6". We established a coexpression network of dysregulated miRNAs and mRNAs to predict the function of miRNAs by using miRWalk database and Pearson correlation coefficient (PCC) analysis. Cellular experiments were used to validate the results of bioinformatics.
First, 69 common DEGs were obtained from datasets GSE43292 and GSE97210 using the limma package in R. Next, a DEG PPI network was constructed. Functional enrichment analysis of DEGs showed that 11 functional pathways were significantly enriched, such as positive regulation of monocyte chemotaxis. Seven common DE-miRNAs were obtained from the GSE99685 dataset and DE-mRNAs predicted miRNAs through the miRWalk database. The miRNA-mRNA network constructed using Cytoscape software suggested that targeted contactin 4 (). Quantitative real-time polymerase chain reaction (qRT-PCR) assay results indicated that was downregulated and was upregulated in the THP-1 + phorbol 12-myristate 13-acetate (PMA) + oxidized low-density lipoprotein (oxLDL) group compared with the THP-1 + PMA group. qRT-PCR, flow cytometry, and enzyme-linked immunosorbent assay (ELISA) found that upregulated significantly inhibited the expression of , cell apoptosis, and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) concentrations in oxLDL-induced THP-1 macrophages. In addition, a dual-luciferase reporter assay demonstrated that was a target gene of .
Overall, these findings suggested that inhibited oxLDL-induced cell apoptosis and inflammation via targeting in THP-1 macrophages.
动脉粥样硬化（AS）严重影响人类健康。微小RNA（miRNA）在AS发病机制和进展中的作用已成为研究热点。我们的目标是确定AS中差异表达的miRNA（DE-miRNA）的生物学效应。
利用基因表达综合数据库（GEO）和limma软件包分析AS中差异表达基因（DEG），包括差异表达的信使核糖核酸（DE-mRNA）和DE-miRNA。通过检索相互作用基因/蛋白的搜索工具（STRING）数据库、Cytoscape软件和Cytoscape插件“ClueGO2.5.6”构建DEG的蛋白质-蛋白质相互作用（PPI）网络并进行功能富集分析。利用miRWalk数据库和皮尔逊相关系数（PCC）分析建立失调miRNA和mRNA的共表达网络以预测miRNA的功能。通过细胞实验验证生物信息学结果。
首先，使用R语言中的limma软件包从数据集GSE43292和GSE97210中获得69个常见的DEG。接下来，构建了DEG的PPI网络。DEG的功能富集分析表明有11条功能通路显著富集，如单核细胞趋化的正调控。从GSE99685数据集中获得7个常见的DE-miRNA，并通过miRWalk数据库由DE-mRNA预测miRNA。使用Cytoscape软件构建的miRNA-mRNA网络表明靶向接触蛋白4（）。定量实时聚合酶链反应（qRT-PCR）检测结果表明，与THP-1+佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）组相比，THP-1+PMA+氧化型低密度脂蛋白（oxLDL）组中（）下调而（）上调。qRT-PCR、流式细胞术和酶联免疫吸附测定（ELISA）发现，上调（）可显著抑制oxLDL诱导的THP-1巨噬细胞中（）的表达、细胞凋亡以及白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的浓度。此外，双荧光素酶报告基因检测表明（）是（）的靶基因。
总体而言，这些发现表明（）通过靶向THP-1巨噬细胞中的（）抑制oxLDL诱导的细胞凋亡和炎症。
