高通量测序筛选冠心病血瘀证相关lncRNA-miRNA-mRNA调控网络

发布时间：2019-01-19 10:17

【摘要】：目的:筛选冠心病血瘀证相关差异表达非编码RNA(lncRNA),微小RNA(microRNA,miRNA),信使RNA(mRNA),构建基因间调控网络,从转录组层面研究冠心病血瘀证物质基础和病理机制。方法:使用高通量测序技术,分别检测冠心病血瘀证、冠心病非血瘀证和正常(各5例)lncRNA,miRNA和mRNA表达情况,通过交联分析筛选冠心病血瘀证相关的差异基因表达谱。对获得的差异基因进行功能通路分析,根据基因间Pearson相关分析和star Base靶基因预测平台,构建基因间调控网络,通过网络拓扑分析,筛选其中的关键基因。在另一匹配队列中(每组各15例)对基因调控网络中的关键节点进行实时荧光定量PCR(Real-time PCR)验证。结果:39个lncRNA,229个miRNA和221个mRNA与冠心病血瘀证密切相关。功能与通路分析结果显示,冠心病血瘀证差异表达基因主要与免疫和炎症相关。共有9个lncRNA(均为下调),31个mRNA(11个上调,20个下调)和24个miRNA(14个上调,10个下调)构成共调控网络,包括76个基因间调控关系。CTA-384D8.35,CTB-114C7.4,RP11-567M16.6和hsa-miR-3158-3p是冠心病血瘀证基因调控网络中的关键节点。Real-time PCR结果验证了上述结果。结论:冠心病血瘀证存在lncRNA-miRNA-mRNA差异表达基因谱,其与免疫和炎症密切相关;差异表达基因间存在相互调控关系,并可依此构建冠心病血瘀证相关lncRNA-miRNA-mRNA调控网络。对冠心病血瘀证的物质基础进行深度挖掘,可为从转录组层面开展中医证型相关研究提供了一定的科学基础。
[Abstract]:Objective: to screen the differential expression of non-coding RNA (lncRNA), microRNA (microRNA,miRNA) in blood stasis syndrome of coronary heart disease (CHD), construct intergene regulatory network of messenger RNA (mRNA), and study the material basis and pathological mechanism of blood stasis syndrome in coronary heart disease (CHD) from the transcriptional level. Methods: high throughput sequencing technique was used to detect the expression of lncRNA,miRNA and mRNA in patients with coronary heart disease (CHD) with blood stasis syndrome, non-blood stasis syndrome and normal (5 cases each). The differentially expressed gene profiles related to blood stasis syndrome of coronary heart disease (CHD) were screened by cross-linking analysis. According to the Pearson correlation analysis between genes and the star Base target gene prediction platform, the intergene regulatory network was constructed, and the key genes were screened by network topology analysis. In another matching cohort (15 cases each), real-time fluorescence quantitative PCR (Real-time PCR) was performed to verify the key nodes in the gene regulatory network. Results: 229 miRNA and 221 mRNA of 39 lncRNA, were closely related to blood stasis syndrome of coronary heart disease. The results of functional and pathway analysis showed that the differentially expressed genes were mainly related to immunity and inflammation in patients with coronary heart disease. A total of 9 lncRNA (all down-regulated), 31 mRNA (11 up-regulated, 20 down-regulated) and 24 miRNA (14 up-regulated, 10 down-regulated) constitute a co-regulatory network, including 76 gene regulatory relationships. RP11-567M16.6 and hsa-miR-3158-3p are the key nodes in the gene regulatory network of coronary heart disease with blood stasis syndrome. Real-time PCR results verify the above results. Conclusion: there is a differential expression gene profile of lncRNA-miRNA-mRNA in blood stasis syndrome of coronary heart disease, which is closely related to immunity and inflammation, and there is a mutual regulation relationship between the differentially expressed genes, which can be used to construct the lncRNA-miRNA-mRNA regulatory network related to blood stasis syndrome of coronary heart disease. The deep excavation of the material basis of the blood stasis syndrome of coronary heart disease can provide a certain scientific basis for the research of TCM syndromes from the transcriptional level.
【作者单位】：中国中医科学院广安门医院;北京中医药大学;
【基金】：国家自然科学基金项目(81673847)
【分类号】：R259

【相似文献】