黄连素作用鼠疫耶尔森菌的体外表达谱

目的 探讨黄连索抑制鼠疫耶尔森菌(简称鼠疫菌)作用的分子机制.方法 应用鼠疫菌全基因组芯片表达谱进行黄连素作用鼠疫菌研究.液体稀释法测定黄连索作用鼠疫菌的最小抑菌浓度(MIC).应用10倍MIC的黄连索作用于鼠疫菌30 min后,提取鼠疫菌总RNA,反转录合成eDNA,Cy3,Cy5荧光染料标记,芯片杂交,扫描仪扫描,SAM软件分析结果 .结果 在鼠疫菌全基因组范围内黄连素作用鼠疫菌的明显差异表达基因有360个.其中明显上调基因333个,下调基因27个.依据鼠疫菌CO92株基冈功能分类结果 ,这些差异基因分属24个不同的功能类群,其中主要有细胞膜相关基冈83个,未知功能基因75个,转运结合蛋白基因48个.发生明显改变的基因功能类群是代谢相关基因,共有40个.结论 获得了黄连素作用鼠疫菌的表达谱.阐述了黄连作用鼠疫菌的分子机制,主要机制在于影响代谢相关基因的上调.

Global gene expression of berberine against Yersiniapestis in vitro

Objective To investigate the antibacterial molecular mechanism of Traditional Chinese Medicine Coptis rhizome against Yersinia pestis(Y.pestis).Methods The method based on whole genome DNA micrnarray of Y.pestis was used.The minimal inhibition concentration(MIC)of berberine to Y.pestis was determined with liquid dilution method.Then gene expression profile of Y.pestis was performed after exposed to berberine at the concentration of 10×MIC for 30 minutes.Total RNA extracted and purified from Y.pestis and reverse-transcribed to cDNA,then labeled by Cy-dye.Finally,the labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software.Results The gene expression profile data revealed that the response of Y.pestis to berberine was a global phenomenon.A total of 360 genes changed significantly.Among them,333 genes were up-regulated,27 down-regulated.These differentially expressed genes were further classified into 24 different functional categories based on the genomie annotation of Y.pestis CO92,in which the number of mainly related genes were 83,75 and 48,including cell envelop,unkown,transport/binding proteins functions.The 40 genes related to the metabolism were upregulated,which was a remarkable change.Conclusion Our results have revealed the general gene expression changes of Y.pestis in response to berberine and demonstrated the antibacterial molecular mechanism of the Coptis rhizome.The major mechanism of Y.pestis in response to berberine is the upregulation of genes related to the metabolism.