血瘀体质中lncRNA和mRNA表达谱分析及调控网络研究

Study on the lncRNA and mRNA Expression Profiles and Regulatory Network in Blood Stasis Constitution

目的:为了更客观地探索血瘀体质的微观特征,本研究从转录组学水平首次初步揭示血瘀体质外周血中lncRNA和mRNA的表达谱特征及其相关作用网络。方法:运用Agilent Human lncRNA+mRNA 4×180 k表达谱芯片,从血瘀体质(BSC,n=11)和平和体质(BC,n=9)者的外周血中分离单个核细胞(Peripheral Blood Mononuclear Cells,PBMCs),采用“limma”方法筛选出2组差异有统计学意义的lncRNAs和mRNAs,对差异基因进行GO功能和KEGG信号通路的显著性富集分析,通过构建蛋白质-蛋白质相互作用(PPI)网络图并寻找其作用通路中关键的hub基因。结果:差异表达的lncRNAs和mRNAs均能够将血瘀体质和平和体质进行区分。血瘀体质中差异有统计学意义的lncRNAs 394个(139个上调,255个下调)和mRNAs 410个(159个上调,251个下调)。结果显示,血瘀体质差异基因显著聚集在生长发育调节和细胞信号传导相关作用通路,例如,MAP激酶活性的正向调控、血管生成的正向调控、腺苷酸环化酶活性的激活、卵巢卵泡发育、γ-氨基丁酸能突触以及PI3K-AKT信号通路等。通过构建PPI并筛选了一些核心基因,如,FGF2、VEGFA、HIST1H2BK和PHF5A等。结论:血瘀体质易感冠心病,脑卒中等循环系统疾病和疼痛性疾病等物质基础可能与lncRNAs/mRNAs的生长发育调节和细胞信号传导失调相关。

Objective:To objectively explore the microscopic characteristics of blood stasis constitution.This study reveals for the first time the expression profile of lncRNA and mRNA in peripheral blood of blood stasis constitution and its related action network from the level of transcriptomics.Methods:lncRNA+mRNA Human Gene Expression Microarray V4.0(4×180 k)was performed in this study.The peripheral blood mononuclear cells(PBMCs)were isolated from subjects between the blood stasis constitution(n=11)and the balanced constitution(n=9).The“limma”method was used to screen out the 2 groups of lncRNAs and mRNAs with statistically significant differences,and we perform the significant enrichment analysis of the GO function and KEGG pathway for the difference genes.By constructing a protein-protein interaction(PPI)network diagram,we found the key hub gene in its action pathway.Results:The differential expression lncRNAs/mRNAs could be used to distinguish individuals with the blood stasis constitution from those with the balanced constitution.These were 394 DElncRNAs of statistically significant(139 up-regulated and 255 down-regulated)and 410 DElncRNAs(159 up-regulated and 251 down-regulated)in the blood stasis constitution.The results indicated that the differences genes in blood stasis constitution were significantly clustered in the pathways related to the dysregulation of growth and development and cell signaling,for example,positive regulation of MAP kinase activity,positive regulation of angiogenesis,activation of adenylate cyclase activity,ovarian follicle development,ovarian follicle development,gamma-aminobutyric acid synapse,and PI3K-AKT signaling pathway,etc.Core genes such as FGF2,VEGFA,HIST1H2BK and PHF5A were screened by constructing a protein-protein interaction network(PPI).Conclusion:Blood stasis constitution is susceptible to coronary heart disease,circulatory system diseases such as stroke and painful diseases.The material basis may be related to the growth and development regulation of lncRNAs/mRNAs and the imbalance of cell signal conduction.