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

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 this study was to screen out the potential candidate microRNAs (miRNAs) in DM patients with BSS by high-throughput sequencing (HTS) and bioinformatics analysis. CRL-1730 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) and mRNAs (DE-mRNAs) were screened between samples. 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. Overall design: Bioinformatics analysis of miRNAs in DM patients with BSS by establishing cell models of DM with BSS.

在传统中医学（Traditional Chinese Medicine, TCM）中，血瘀证（blood stasis syndrome, BSS）主要表现为血液黏度、血小板黏附率与聚集率升高，以及微循环改变，进而导致血管内皮损伤，是糖尿病（diabetes mellitus, DM）发生发展的重要危险因素。本研究旨在通过高通量测序（high-throughput sequencing, HTS）与生物信息学分析，筛选伴血瘀证糖尿病患者的潜在候选微小核糖核酸（microRNAs, miRNAs）。将CRL-1730株人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVECs）与10%人血清共孵育，构建伴血瘀证糖尿病（DM+BSS组）、不伴血瘀证糖尿病（NBS组）及正常对照（NC组）细胞模型。提取各组样本的总RNA，采用Hiseq2000测序平台完成测序。筛选各组样本间的差异表达微小核糖核酸（Differentially expressed miRNAs, DE-miRNAs）与差异表达信使核糖核酸（Differentially expressed mRNAs, DE-mRNAs）。通过生物信息学软件预测微小核糖核酸的靶基因，并对靶基因开展基因本体（Gene Ontology, GO）富集分析与通路富集分析。基于P值≤0.001的显著富集基因本体注释与通路，筛选伴血瘀证糖尿病的关键微小核糖核酸。整体实验设计：通过构建伴血瘀证糖尿病细胞模型，对伴血瘀证糖尿病患者的微小核糖核酸开展生物信息学分析。