大黄抑制鼠疫耶尔森菌的体外转录组学研究

The transcriptomics study on the antibacterial mechanism of rubharb against Yersinia pestis in vitro

目的应用鼠疫菌全基因组芯片研究大黄抑制鼠疫菌的分子作用机制。方法液体稀释法测定大黄对鼠疫菌的最低抑菌浓度(MIC),以10倍MIC作用鼠疫菌30min,提取鼠疫菌总RNA,逆转录合成cDNA,荧光素标记,与芯片杂交,扫描仪扫描,应用SAM软件分析结果。结果获得了大黄作用鼠疫菌的表达谱。大黄作用鼠疫菌的明显差异基因为498个,其中上调基因358个,下调140个。结论蛋白质合成基因、细胞膜相关基因、转运结合蛋白基因以及部分热休克基因的改变是大黄抑制鼠疫菌的主要作用机制。

The molecular mechanism of the antibacterial action of the Chinese traditional medicine rubharb against Yersinia pestis was investigated in vitro, in which the method of DNA microarray based on gene expression profile was used. The minimal inhibition eoncentration（MIC） of rubharb against Y. Destis was determined with liquid dilution method and the gene expression profile of this organism was performed after exposure to rubharb at concentration of 10 MIC for 30 minutes. Meanwhile, the total RNA was extracted and purified from Y. pestis, reversely transcribed to eDNA and then labeled by Cydye. Finally, the labeled probes were hybridized to microarray and the results were demonstrated by a laser scanner. The results were analyzed by SAM software. In these ways, the expression profile of rubharb against Y. pestis could be obtained and the transcriptional profile data revealed that the response of Y. pestis to rubharb was a global phenomenon. As a whole, 498 genes were altered significantly, among which 358 genes were up-regulated and 140 genes down-regulated. From these observak tion, it seems that the mechanism of the main action of rubharb against Y. pestis would be alterations on the genes related to protein synthesis ,cell envelope-associated gene, transport-binding protein gene and certain heat shock protein genes.