利血平对化脓隐秘杆菌大环内酯类外排基因mefA表达的影响  

Effect of reserpine on the expression of mefA in T.pyogenes

作　　者：苏鸿雨 黄程程[1] 刘耀川[1] 陈梦涵 朱晓明[1] 刘利鹏 张德显[1] 刘明春[1] SU Hongyu;HUAGN Chengcheng;LIU Yaochuan;CHEN Menghan;ZHU Xiaoming;LIU Lipeng;ZHANG Dexian;LIU Mingchun(College of Animal Husbandry and Veterinary Medicine,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]沈阳农业大学,畜牧兽医学院,辽宁沈阳110866

出　　处：《中国兽医学报》2021年第1期91-95,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31572564,31972736)。

摘　　要：化脓隐秘杆菌(Trueperella pyogenes,T.pyogenes)是一种能够引起动物和人化脓性感染的重要病原菌。随着抗生素的广泛使用,该菌已对临床常用的大环内酯类、四环素类等药物产生不同程度的耐药性。本试验拟探讨外排泵抑制剂利血平对T.pyogenes大环内酯类外排基因mefA mRNA及蛋白表达的影响,将为细菌外排泵抑制剂的研究奠定理论基础。以携带mefA基因的T.pyogenes分离株(17,20号)为试验菌株,采用荧光定量PCR及Western blot技术,检测利血平作用前后各菌株mefA基因mRNA及蛋白表达量。结果表明,1/2 MIC利血平作用2株耐药菌株36 h后,大环内酯类抗生素红霉素、罗红霉素、泰乐菌素、阿奇霉素和替米考星对携带mefA基因的T.pyogenes的MIC值均有不同程度的降低。17和20号分离株外排基因mefA mRNA表达水平较药物作用前均极显著降低(利血平作用前是作用后的5.18,17.54倍),外排蛋白MefA表达量较药物作用前也显著降低(利血平作用前是作用后的1.08,1.70倍),且基因水平和蛋白水平的变化与其耐药表型的变化趋势一致;提示利血平可能是通过抑制外排基因mefA的转录和翻译而消减T.pyogenes对大环内酯类抗生素的耐药性。Trueperella pyogenes(T.pyogenes)is an important pathogen.It causes different levels of suppurative infection in various animals because of abuse and unreasonable use of antibiotics,especially macrolides and tetracyclines.mefA gene plays an important role in the regulation of macrolide resistance.The purpose of this study was to explore the effect of reservoir element on the expression of T.pyogenes mefA gene,and to provide a theoretical basis for the study of bacterial excretory pump inhibitors.Two strains(No.17 and 20)of T.pyogenes,as a mefA gene vector,were used as the test strains.Quantitative fluorescent PCR and Western blot were used to determine the mefA gene expression level of each strain before and after the treatment of resperine.The results showed that after 1/2 MIC of reserpine treatment on the resistant strains for 36 h,the macrolide antibiotics(including erythromycin,spiramycin,tyrosine,azithromycin and temiclocine)reduced the MIC value of T.pyogenes carrying mefA gene to varying degrees.The mefA mRNA levels of two strains decreased by 5.18 times and 17.54 times respectively compared with that before the drug treatment.The protein MefA decreased by 1.08 times and 1.70 times respectively compared with that before the drug treatment.The changes of mRNA and protein levels were consistent with the changes of drug resistance phenotypes.Storage protein can inhibit the transcription and translation of mefAforeign gene,thus reducing the resistance of T.pyogenes to macrolides.

关 键 词：化脓隐秘杆菌 利血平 外排基因mefA 外排泵抑制剂 

分 类 号：S852.61[农业科学—基础兽医学] S859.796[农业科学—兽医学]