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首页> 美国卫生研究院文献>PLoS Clinical Trials >Discovery of Molecular Mechanisms of Traditional Chinese Medicinal Formula Si-Wu-Tang Using Gene Expression Microarray and Connectivity Map
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Discovery of Molecular Mechanisms of Traditional Chinese Medicinal Formula Si-Wu-Tang Using Gene Expression Microarray and Connectivity Map

机译：基因表达芯片和连接图谱发现中药四物汤分子机理

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To pursue a systematic approach to discovery of mechanisms of action of traditional Chinese medicine (TCM), we used microarrays, bioinformatics and the “Connectivity Map” (CMAP) to examine TCM-induced changes in gene expression. We demonstrated that this approach can be used to elucidate new molecular targets using a model TCM herbal formula Si-Wu-Tang (SWT) which is widely used for women's health. The human breast cancer MCF-7 cells treated with 0.1 µM estradiol or 2.56 mg/ml of SWT showed dramatic gene expression changes, while no significant change was detected for ferulic acid, a known bioactive compound of SWT. Pathway analysis using differentially expressed genes related to the treatment effect identified that expression of genes in the nuclear factor erythroid 2-related factor 2 (Nrf2) cytoprotective pathway was most significantly affected by SWT, but not by estradiol or ferulic acid. The Nrf2-regulated genes HMOX1, GCLC, GCLM, SLC7A11 and NQO1 were upreguated by SWT in a dose-dependent manner, which was validated by real-time RT-PCR. Consistently, treatment with SWT and its four herbal ingredients resulted in an increased antioxidant response element (ARE)-luciferase reporter activity in MCF-7 and HEK293 cells. Furthermore, the gene expression profile of differentially expressed genes related to SWT treatment was used to compare with those of 1,309 compounds in the CMAP database. The CMAP profiles of estradiol-treated MCF-7 cells showed an excellent match with SWT treatment, consistent with SWT's widely claimed use for women's diseases and indicating a phytoestrogenic effect. The CMAP profiles of chemopreventive agents withaferin A and resveratrol also showed high similarity to the profiles of SWT. This study identified SWT as an Nrf2 activator and phytoestrogen, suggesting its use as a nontoxic chemopreventive agent, and demonstrated the feasibility of combining microarray gene expression profiling with CMAP mining to discover mechanisms of actions and to identify new health benefits of TCMs.
机译：为了寻求系统的方法来发现中药（TCM）的作用机理，我们使用了微阵列，生物信息学和“连接图”（CMAP）来检查中药诱导的基因表达变化。我们证明了这种方法可用于使用模型TCM草药配方Si-Wu-Tang（SWT）阐明新的分子靶标，该模型广泛用于女性健康。用0.1 µM雌二醇或2.56 mg / ml SWT处理的人乳腺癌MCF-7细胞显示出显着的基因表达变化，而未发现阿魏酸（一种已知的SWT生物活性化合物）发生显着变化。使用与治疗效果相关的差异表达基因进行的途径分析表明，核因子红系2相关因子2（Nrf2）细胞保护途径中的基因表达受SWT的影响最大，而不受雌二醇或阿魏酸的影响。 SWT以剂量依赖的方式上调了Nrf2调控的基因HMOX1，GCLC，GCLM，SLC7A11和NQO1，并通过实时RT-PCR进行了验证。一致地，用SWT及其四种草药成分进行处理后，MCF-7和HEK293细胞中的抗氧化反应元件（ARE）-萤光素酶报道分子活性增加。此外，将与SWT处理相关的差异表达基因的基因表达谱与CMAP数据库中的1,309种化合物进行了比较。雌二醇处理的MCF-7细胞的CMAP图谱显示与SWT处理非常匹配，这与SWT广泛用于妇女疾病的用途一致，并表明具有植物雌激素作用。化学预防剂与铁传递素A和白藜芦醇的CMAP图谱也显示出与SWT的图谱高度相似。这项研究将SWT鉴定为Nrf2激活剂和植物雌激素，表明其可作为无毒化学预防剂使用，并证明了将微阵列基因表达谱与CMAP挖掘相结合以发现其作用机制并确定中医对健康的新益处的可行性。

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