高等学校化学学报 ›› 2012, Vol. 33 ›› Issue (11): 2397.doi: 10.7503/cjcu20120347

• 分析化学 • 上一篇    下一篇

基因芯片用于组分中药新双龙方的配伍机制研究

吕琳星1, 范雪梅2, 梁琼麟2, 王文凯1, 王义明1,2, 罗国安1,2   

  1. 1. 江西中医学院药学院, 南昌 330004;
    2. 清华大学化学系, 北京 100084
  • 收稿日期:2012-04-13 出版日期:2012-11-10 发布日期:2012-10-15
  • 通讯作者: 罗国安,男,教授,博士生导师,主要从事生命分析化学相关研究.E-mail:luoga@mail.tsinghua.edu.cn E-mail:luoga@mail.tsinghua.edu.cn
  • 基金资助:

    国家自然科学基金重点项目(批准号: 81130066)和科技部国际合作项目(批准号: 2010DFA32420)资助.

cDNA Microarray for the Study of Compatibility Mechanism of Multi-component Chinese Medicine New Shuanglong Formula

LÜ Lin-Xing1, FAN Xue-Mei2, LIANG Qiong-Lin2, WANG Wen-Kai1, WANG Yi-Ming1,2, LUO Guo-An1,2   

  1. 1. School of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China;
    2. Department of Chemistry, Tsinghua University, Beijing 100084, China
  • Received:2012-04-13 Online:2012-11-10 Published:2012-10-15

摘要:

采用基因芯片技术研究了中药新双龙方及其有效组分人参总皂苷和丹参总酚酸治疗急性心肌梗塞(AMI)大鼠的作用机制. 以结扎大鼠冠脉左前降支的方法制备急性心梗模型, 采用大鼠全基因组表达谱芯片分别筛选出AMI大鼠给药前及新双龙方与组分给药后的差异表达基因(Ratio>2或<0.5), 通过对4组差异基因的综合分析, 比较了新双龙方与组分作用方式的异同, 从分子调控水平上探讨复方配伍后的增效作用, 同时对与药效作用机理直接相关的钙离子转运相关的Cacna1d基因和血管修复相关的Flnb基因进行了实时荧光逆转录聚合酶链式反应(RT-PCR)定量. 结果表明, 新双龙方与2种有效组分对心梗大鼠基因表达的影响不同. 对差异基因的聚类分析以及药物的调控作用比较结果均表明复方比组分的疗效更好; 通路分析结果表明, 新双龙方与组分均主要通过调控钙信号转导通路、 丝裂原活化蛋白激酶信号转导通路(MAPK)信号等通路发挥药效作用, 且复方涉及更多通路; 筛选出了复方与组分共同调控的靶点基因, 其在3组中具有不完全相同的表达趋势, 但复方具有更积极的调控作用. 本研究结果从分子调控角度证实了复方新双龙方的配伍优势.

关键词: 基因表达谱, 实时荧光定量聚合酶链式反应, 复方配伍, 新双龙方, 急性心肌梗塞

Abstract:

The pharmacological mechanism of New Shuanglong Formula with its two effective components total ginsenoside(TGS) and total salvianolic acid(TSA) on acute myocardial infarction(AMI) rats was compared by cDNA microarray. The model was induced by ligation of the left anterior descending artery, and significant genes were screened(Ratio>2 or <0.5) when AMI rats were treated before and after with New Shuanglong Formula and components by genome-wide expression profiles. Through comprehensive analysis of significant genes of four groups, the similarities and differences of effect between New Shuanglong Formula and components were analyzed and the synergy of compound compatibility at molecular control level was discussed; meanwhile, quantitative real time reverse transcription-polymerase chain reaction(RT-PCR) was used to analyse Cacna1d gene(associated with Calcium ion transport)and Flnb gene(associated with Vascular repair) which both were directly related to the pharmacological effects. The analysis of significant genes showed the effect on gene expression of AMI rats was different from New Shuanglong Formula to components, the compound appeared to play a better effect than components. Through clustering analysis of different genes and the comparison of regulation effects by drugs, the compound appeared to play a better effect than componects. Pathway analysis indicates the affects of New Shuanglong Formula and components are mainly through regulating calcium signaling pathway, MAPK signaling pathway and so on, and the compound involved more pathways; common target genes which are regulated by compound and components were screened, although their expression patterns are not exactly the same, but the compound appear to play a better effect. The results of this study have illuminated the advantage of New Shuanglong Formula from the perspective of molecular control.

Key words: Gene expression profile, Real time reverse transcription-polymerase chain reaction(RT-PCR), Compatibility of Traditional Chinese Medicine(TCM), New Shuanglong Formula, Acute Myocardial infarction

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