基于基因芯片技术的咳喘六味合剂治疗寒哮型支气管哮喘的靶点发现和机制研究

The target-finding and mechanism of drug action researching of Kechuan Liuwei Mixture treating asthma of cold type based on cDNA microarrays technology

关键词:  咳喘六味合剂  寒性哮喘  基因芯片  靶点基因  作用机制

Key words:  Kechuan Liuwei Mixture  asthma of cold type  cDNA microarrays technology  target gene  mechanism of drug action

摘要:

[目的] 利用基因芯片技术探索咳喘六味合剂治疗寒哮型支气管哮喘作用的基因靶点及相关机制。[方法] 选取16例寒哮发作期患者，其中8例服用咳喘六味合剂，8例服用空白对照口服液，服药方法每次20 mL，每日3次，疗程1周。分别于干预后取患者空腹外周血，抽提RNA行基因芯片表达谱差异分析，以基因表达倍数值1.5和基因表达倍数值-1.5为阈值来确定差异表达基因，然后用Gene Ontology及Pathway分析对差异表达基因进行功能分类及生物信号通路分析。[结果] 通过基因芯片技术，筛选出差异表达1.5倍以上的基因有233条，其中上调基因106条，下调基因127条。咳喘六味合剂治疗寒性哮喘过程中差异表达的基因集中在免疫、炎症、血管生成负调节、肿瘤坏死因子受体连接功能区和细胞因子-受体相互作用、黏着斑、钙信号、细胞外基质受体相互作用等生物学通路。[结论] 咳喘六味合剂治疗寒哮可能通过干预与免疫、炎症、气道重塑、钙离子通道等相关的基因的表达从多途径、多靶点发挥作用。

Abstract:

[Objective] The aim of this study was to explore the target genes and mechanism of drug action of Kechuan Liuwei Mixture.[Methods] The 16 patients who were diagnosed with asthma of cold type were selected to do the gene chip screening. The 8 cases of them took Kechuan Liuwei Mixture three times a day, 20 mL per time, while the others took the placebo. Both groups were treated for one week. Peripheral blood samples were taken after one-week drug intervention. Total RNA was extracted from peripheral blood. Using gene expression times numerical 1.5 and -1.5 for threshold to determine the differential expression genes,then GO and pathway analysis were used to analyze the differential expression genes on function and classification.[Results] Screening and identification of differentially expressed genes, 233 1.5-fold DEGs were screened from the gene expression spectrum, while 106 genes were up-regulated and 127 genes were down-regulated. These differently expressed genes are involved in functions of inflammatory responses,immune response,Cytokine-cytokine receptor interaction,transforming growth factor and pathways of Cytokine-cytokine receptor interaction, Focal adhesion, Calcium signaling pathway, MAPK signaling pathway, ECM-receptor interaction of asthma.[Conclusion] Kechuan Liuwei Mixture could treat asthma of cold type through multi-channels and multi-targets by effecting genes that related to immune response, inflammatory reaction, airway remodeling and calcium channels.