冠心病血瘀证转录组学研究——病证结合生物标志物研究思路与方法

发布时间：2018-03-24 09:27

本文选题：冠心病　切入点：血瘀证　出处：《中国实验方剂学杂志》2017年19期

【摘要】：中医证候的生物学基础研究是影响中医学发展与创新的重大科学问题,证候相关生物标志物及中药干预调控网络的研究对于在证候标准规范的基础上进一步提高诊断效能和临床疗效具有重要意义。因此,在前期冠心病证候标准规范研究的基础上,本项目组以冠心病心绞痛血瘀证为切入点,从分子生物水平对证候进行了深入研究。第一,通过采集累计312例患者外周血基因信息,运用差异显示法、实时定量聚合酶链式反应(Real-time polymerase chain reaction,Real-time PCR),斑点杂交法,高通量芯片,测序等方法,确立了冠心病血瘀证长链非编码核糖核酸-微小核糖核酸-信使核糖核酸(long noncoding ribonucleic acid-micro ribonucleic acid-messenger ribonucleic acid,lncRNA-miRNA-mRNA)3个层面的潜在生物标志物。第二,在前期筛选出的差异基因的基础上,运用生物信息学分析结合细胞功能学验证的方法,构建了冠心病血瘀证基因miRNAmRNA以及lncRNA-miRNA-mRNA 2个水平的调控网络。第三,基于血瘀证差异基因及调控网络,通过累计202例活血化瘀中药干预冠心病心绞痛血瘀证患者的随机对照试验,治疗前后共计400余人次的基因检测,以及活血化瘀中药处理相关细胞模型的体外实验,从基因-蛋白-功能3个层面研究中药作用机制,从分子水平揭示了活血化瘀中药干预冠心病血瘀证的基因调控网络机制。本研究将相关的分子生物学技术及生物信息学方法引入证候的生物学基础研究,建立了一套证候相关生物标志物的研究方法,为证候实质的研究提供了新思路。
[Abstract]:The biological basic research of TCM syndromes is an important scientific problem affecting the development and innovation of TCM. The study of syndrome-related biomarkers and TCM intervention and control network is of great significance for further improving the diagnostic efficacy and clinical efficacy on the basis of the criteria of syndromes. Therefore, on the basis of the study on the standard norms of coronary heart disease syndrome in the early stage, This project group took angina pectoris and blood stasis syndrome of coronary heart disease as the breakthrough point, carried on the thorough research from the molecular biological level to the syndrome. First, through collecting the cumulative 312 patients' peripheral blood gene information, the differential display method was used. Real-time polymerase chain reactionation real-time PCR, dot blot hybridization, high-throughput microarray, sequencing, etc. The potential biomarkers of long noncoding ribonucleic acid-micro ribonucleic acid-messenger ribonucleic ribonucleic lncRNA-miRNA-mRNA-mRNA-mRNAs in long chain non-coding ribonucleic acids (RNs) and microRNA (RNs) in patients with coronary heart disease (CHD) with blood stasis syndrome were established. Second, based on the differential genes screened in the early stage, the potential biomarkers of long noncoding ribonucleic acid-micro ribonucleic acid-messenger ribonucleic acidincRNA-mRNAs were established. With the method of bioinformatics analysis and cell function verification, the regulatory networks of miRNAmRNA and lncRNA-miRNA-mRNA were constructed. Thirdly, based on the differential genes and regulatory networks of blood stasis syndrome, A randomized controlled trial was conducted in 202 patients with angina pectoris and blood stasis syndrome of coronary heart disease treated with traditional Chinese medicine for promoting blood circulation and removing blood stasis. The gene detection of more than 400 people before and after treatment and the in vitro experiment of treating the related cell model with traditional Chinese medicine for promoting blood circulation and removing blood stasis were carried out. To study the action mechanism of traditional Chinese medicine from three aspects of gene, protein and function. The mechanism of gene regulation network of traditional Chinese medicine for promoting blood circulation and removing blood stasis in patients with coronary heart disease (CHD) was revealed from molecular level. In this study, molecular biological techniques and bioinformatics methods were introduced into the biological basic study of syndrome. A set of research methods of syndromes related biomarkers is established, which provides a new idea for the study of syndromes essence.
【作者单位】：中国中医科学院广安门医院;北京中医药大学东直门医院;首都医科大学附属北京中医医院;北京中医药大学;
【基金】：国家自然科学基金项目(81473561,81673847)
【分类号】：R259

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