鱼腥草素钠及其与红霉素联用对产膜表皮葡萄球菌luxS,agr/RNAⅢ影响的研究  

Effect of sodium houttuyfonate in combination with erythromycin on luxS,agr/RNAⅢ of Staphylococus epidermidis

作　　者：许甘霏[1] 王晶晶[1] 吴大强[2] 官妍[2] 

机构地区：[1]安徽大学生命科学学院,安徽合肥230601 [2]安徽中医药大学中西医结合临床学院,安徽合肥230038

出　　处：《中国中药杂志》2017年第11期2131-2138,共8页China Journal of Chinese Materia Medica

基　　金：安徽省自然科学基金项目(1508085MH200);安徽省教育厅自然科学研究项目(KJ2013Z163)

摘　　要：细菌的群体感应及特异基因表达调控在细菌生物被膜形成中有十分重要的作用。luxS,agr均是表皮葡萄球菌群体感应调控中的关键基因,RNAⅢ是agr系统的效应器分子。为了评估鱼腥草素钠及其与红霉素联用对产膜表皮葡萄球菌转录水平的影响,采用连续稀释法测定鱼腥草素钠、红霉素、万古霉素对表皮葡萄球菌的MIC;荧光定量PCR测定鱼腥草素钠及其与红霉素联用、万古霉素、红霉素等对表皮葡萄球菌作用后不同时间段luxS,agr/RNAⅢ的转录水平。结果表明1/2MIC,1/4MIC鱼腥草素钠,1/2MIC鱼腥草素钠+1/2MIC红霉素,1/4MIC鱼腥草素钠+1/4MIC红霉素,1/8MIC鱼腥草素钠+1/8MIC红霉素等在作用于ATCC 35984伊始,就可迅速上调luxS的表达,与阴性对照相比,存在显著差异(P<0.05);在作用6,12,48h后,鱼腥草素钠依然存在对luxS的上调作用。鱼腥草素钠在MIC,1/2MIC浓度时对agr表达均有显著下调作用(P<0.05),1/2MIC鱼腥草素钠与1/2MIC红霉素联用、1/4 MIC鱼腥草素钠与1/4MIC红霉素联用,在作用6,12,24 h等时间段对agr表达的下调作用也很显著(P<0.05)。鱼腥草素钠在MIC浓度时,对RNAⅢ表达亦有显著下调作用(P<0.05),1/2MIC鱼腥草素钠与1/2MIC红霉素联用对RNAⅢ表达的下调效果也很显著(P<0.05)。提示鱼腥草素钠及其与红霉素联用,可以迅速上调表皮葡萄球菌luxS转录水平的表达,在特定浓度下可下调agr/RNAⅢ的表达,具有抑制表皮葡萄球菌生物被膜菌之间的相互黏附聚集、抑制膜内营养和输水通道的形成、阻止膜内细菌脱离、并抑制细菌外毒素形成等作用的可能。Quorum sensing of bacteria and its specific gene expression regulation have a very important role in bacterial biofilm formation. LuxS and agr are the key regulatory genes in quorum sensing of Staphylococcus epidermidis, and RNA Ⅲ is the effector molecule of agr system. In order to evaluate the effects of sodium houttuyfonate in combination with erythromycin on the transcription level of S. epidermidis, serial dilution method was used to determine the MIC of sodium houttuyfonate, erythromycin and vancomycin on S. epidermidis, and fluorescent quantitative PCR method was used to detect the transcription levels of luxS, agr/RNA m in different time periods after treatment on S. epidermidis by sodium houttuyfonate in combination with erythromycin, vancomycin, and erythromycin alone. Our results showed that in treatment by 1/2MIC, 1/4MIC sodium houttuvfonate, 1/2MIC sodium houttuvfonate ＋ 1/2MIC erythromycin, 1/4MIC sodium houttuyfonate ＋ 1/4MIC erythromycin, and 1/8MIC sodium houttuyfonate ＋ 1/8MIC erythromycin for ATCC 35984, they could rapidly up-regulate the expression of luxS of S. epidermidis from the beginning as compared with negative control, with significant differences （P 〈 0.05） ; furthermore, sodium houttuyfonate can still up-regulate the expression of luxS even after treatment for 6, 12 and 48 h. Sodium houttuyfonate in MIC and 1/2MIC concentration can significantly down-regulate the expression of agr （P 〈 0. 05） ; 1/2MIC sodium houttuyfonate ＋ 1/2MIC erythromycin, 1/4MIC sodium houttuyfonate ＋ 1/4MIC erythromycin, can also significantly down-regulate the expression of agr in 6 h, 12 h and 24 h（ P 〈 0. 05 ）. Sodium houttuyfonate in MIC, can significantly down-regulate the expression of RNA Ⅲ（P 〈 0. 05）, and 1/2MIC sodium houttuyfonate ＋ 1/2MIC erythromycin can also significantly down-regulate the expression of RNA Ⅲ （P 〈 0. 05）. Therefore, our presented results showed that sodium houttuyfonate in combination with erythromycin can rapidly up-regulate the tran

关 键 词：表皮葡萄球菌 鱼腥草素钠 红霉素 群体感应 LUXS agr/RNAⅢ 生物被膜 

分 类 号：R285[医药卫生—中药学]