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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
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国家自然科学基金项目(81473561,81673847)


Transcriptomic Studies on Coronary Heart Disease with Blood Stasis Syndrome——Research Approaches and Methods in Terms of Biomarker and Disease Symptoms
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    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 non-coding 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miRNA-mRNA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:

    Research on biological foundation of traditional Chinese medicine (TCM) syndrome is another critical issue that could affect the development and innovation of Chinese medicine. The studies on syndrome-related biomarker and TCM intervention network have important significance for further enhancing the diagnostic capability and clinical efficacy on the basis of standard syndrome criteria. Hence, based on the previous syndrome standard coronary heart disease (CHD), we started our molecular biology research with blood stasis syndrome (BBS) for coronary heart disease, angina pectoris to deepen the study from molecular biological level. Firstly, we accumulated genetic information from 312 patients and employed differential display, reverse northern, Real-time PCR, spot hybridization, high-throughput chip and sequencing to screen the potential biomarkers of BBS with CHD from long non-coding ribonucleic acid (lncRNA), micro ribonucleic acid (miRNA) and messenger ribonucleic acid (mRNA). Secondly, we constructed the regulating networks of BBS with CHD from miRNA-mRNA and lncRNA-miRNA-mRNA by bioinformatics analysis based on the screened potential biomarkers and then validated the networks by cellular experiment. Thirdly, based on the differential genes and regulating networks mentioned above, we utilized randomized controlled trials enrolling accumulative 202 patients with more than 400 gene tests as well as relevant cellular experiments to explore the action of herbs that promoted blood circulation and removes blood stasis from the aspects of gene, protein and phenotype. The molecular biology techniques and bioinformatics analysis were combined into the biological foundation research of syndrome to establish a series of research methods for syndrome-related biomarkers, providing a new perspective to studying the essence of syndrome.

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王阶,姚魁武,刘咏梅,邢雁伟,李军,杨保林,杨戈,虞桂,滕菲,廖江铨,段练,张振鹏,何庆勇,陈光.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[J].中国实验方剂学杂志,2017,23():1~5

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  • 收稿日期:2017-04-17
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  • 在线发布日期: 2017-08-31
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