Bioinformatics analysis of microRNAs related to blood stasis syndrome in diabetes mellitus patients
Bioinformatics analysis of microRNAs related to blood stasis syndrome in diabetes mellitus patients
复制标题
糖尿病患者血瘀证相关microRNA的生物信息学分析
DOI：
10.1042/bsr20171208
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发表时间：
2018-04-27
影响因子：
4
通讯作者：
Fang, Meixia
中科院分区：
文献类型：
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作者：
Chen, Ruixue;Chen, Minghao;Fang, Meixia
在中医学中，血瘀证主要表现为血液粘度、血小板粘附率和聚集率增高，微循环改变，导致血管内皮损伤。它是糖尿病（DM）发展的重要因素。本研究的目的是通过高通量测序和生物信息学分析筛选出可能与糖尿病合并BSS相关的microRNA（miRNAs）。采用人脐静脉内皮细胞（HUVECs）与10%人血清孵育，建立糖尿病合并BSS、糖尿病不合并BSS（NBS）和正常对照（NC）模型。每个样本提取总RNA，通过Hiseq 2000平台测序。在样品之间筛选差异表达的miRNA（DE-miRNA），并与mRNA丰度的已知变化进行比较。通过软件预测miRNAs的靶基因。对目的基因进行基因本体（GO）和途径富集分析。根据显著富集的GO注释和途径（P值≤ 0.001），我们选择了DM与BSS的关键miRNAs。结果显示，血瘀证患者DE-miRNAs的数量为32个，而非血瘀证患者和正常对照组DE-miRNAs的数量为32个。从GO注释中选择潜在的候选miRNA，其中靶基因显著富集（− log 10（P值）> 5），包括miR-140- 5 p、miR-210、miR-362- 5 p、miR-590- 3 p和miR-671- 3 p。本研究通过HTS和生物信息学分析筛选出与糖尿病合并BSS相关的候选miRNAs。这些miRNAs的发现将有助于在基因水平上为糖尿病合并BSS的临床研究提供有价值的建议。
In traditional Chinese medicine (TCM), blood stasis syndrome (BSS) is mainly manifested by the increase of blood viscosity, platelet adhesion rate and aggregation, and the change of microcirculation, resulting in vascular endothelial injury. It is an important factor in the development of diabetes mellitus (DM). The aim of the present study was to screen out the potential candidate microRNAs (miRNAs) in DM patients with BSS by high-throughput sequencing (HTS) and bioinformatics analysis. Human umbilical vein endothelial cells (HUVECs) were incubated with 10% human serum to establish models of DM with BSS, DM without BSS (NBS), and normal control (NC). Total RNA of each sample was extracted and sequenced by the Hiseq2000 platform. Differentially expressed miRNAs (DE-miRNAs) were screened between samples and compared with known changes in mRNA abundance. Target genes of miRNAs were predicted by softwares. Gene Ontology (GO) and pathway enrichment analysis of the target genes were conducted. According to the significantly enriched GO annotations and pathways (P-value ≤ 0.001), we selected the key miRNAs of DM with BSS. It showed that the number of DE-miRNAs in BSS was 32 compared with non-blood stasis syndrome (NBS) and NC. The potential candidate miRNAs were chosen from GO annotations in which target genes were significantly enriched (−log10 (P-value) > 5), which included miR-140-5p, miR-210, miR-362-5p, miR-590-3p, and miR-671-3p. The present study screened out the potential candidate miRNAs in DM patients with BSS by HTS and bioinformatics analysis. The miRNAs will be helpful to provide valuable suggestions on clinical studies of DM with BSS at the gene level.