AI-2对鸭疫里氏杆菌粘附入侵Vero细胞及其相关基因转录水平的影响  

Effect of autoinducer 2 on Riemerella antatipestifer adherence and invasion to Vero cells

作　　者：刘蕾[1] 韩先干[1] 刘瑞[1] 白灏[1] 董洪亮[1] 丁铲[1] 刘海文[1] 于圣青[1] 

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《微生物学报》2013年第3期313-319,共7页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31072161;31001078)~~

摘　　要：【目的】自诱导信号分子AI-2(Autoinducer-2,AI-2)作为细菌间的通用自诱导信号分子,参与细菌众多生理功能的调控。本研究通过开展AI-2在鸭疫里氏杆菌(Riemerella antatipestifer,RA)CH3株对非洲绿猴肾细胞(Vero细胞)的粘附、入侵以及对RA相关基因调控作用的研究,为进一步研究AI-2对RA的调控作用奠定基础。【方法】在CH3(血清型1型)对Vero细胞的粘附、入侵过程中加入不同浓度的AI-2,研究其对CH3粘附、入侵Vero的影响;在添加184.0μmol/L AI-2的胰蛋白胨大豆肉汤(TSB)中培养RA CH3菌株,利用real-time PCR来检测AI-2对RA相关基因转录水平的影响。【结果】结果表明,AI-2浓度为18.4μmol/L时,AI-2对CH3粘附Vero细胞的抑制性最强,为62%,当AI-2浓度为184.0μmol/L时,AI-2对CH3入侵Vero细胞的促进性最强为194%。荧光定量PCR结果表明AI-2对部分基因的转录有促进作用,对部分基因的转录有抑制作用。【结论】AI-2参与调控RA粘附、入侵Vero细胞及RA毒力因子、免疫原性蛋白基因以及代谢相关基因转录水平的调控。[ Objective ] Autoinducer 2 （AI-2）, used to communicate among bacterial species, regulates numerous physiological functions of bacteria. In this study, we studied the effect of AI-2 on adherence and invasion of Riemerella antatipestifer （RA） strain CH3 to Vero cells and transcriptional levels of virulence-related and metabolism-related genes were investigated. [ Methods] To verify whether the adherence and invasion of CH3 was affected by AI-2, we added different concentrations of AI-2 to the cocuhures of Vero cells and CH3 and then calculated adherence percentages and invasion percentages of tested groups. We further added AI-2 （184.0μmol/L） to the tryptone soya broth culture of CH3 and then detected the effect of transcriptional levels of related genes of CH3 using real-time PCR. [ Results ] The adherence of CH3 to Vero cells was decreased most to 62% with 18.4μmol/L AI-2 and the invasion of CH3 to Vero cells was increased most to 194% with 184. 0μmol/L AI-2. The result of real-time PCR shows that AI-2 increased transcriptional levels of some virulence-related genes and decreased transcriptional levels of some metabolism-related genes. [ Conclusion] These results suggest that AI-2 affected adherence and invasion of CH3 to Vero cells. Moreover, AI-2 could regulate some genes of CH3 to modulate particular physiological behaviors.

关 键 词：鸭疫里氏杆菌 粘附 入侵 非洲绿猴肾细胞(Vero) 自诱导信号分子AI-2 REAL-TIMEPCR 

分 类 号：S852.61[农业科学—基础兽医学]