盐酸白屈菜红碱逆转肺炎克雷伯菌中tmexCD1-toprJ1介导的高水平替加环素抗性  

Chelerythrine chloride reverses high levels of tigecycline resistance mediated by tmexCD1-toprJ1in Klebsiella pneumoniae

作　　者：杨金典 徐蕾 吕冠辰 崔民河 朱先鹏 邓旭明 YANG Jindian;XU Lei;LYU Guanchen;CUI Minhe;ZHU Xianpeng;DENG Xuming(College of Veterinary Medicine,Jilin University,Changchun 130062,China;Jilin Mushuo Breeding Co.,Ltd.,Changchun 130500,China;Agricultural Comprehensive Administrative Law Enforcement Brigade of Liuhe County,Tonghua,Jilin 135300,China)

机构地区：[1]吉林大学动物医学学院,吉林长春130062 [2]吉林省牧硕养殖有限公司,吉林长春130500 [3]吉林省柳河县农业综合行政执法大队,吉林通化135300

出　　处：《中国兽医学报》2024年第4期748-756,共9页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(U22A20523,32202856);中国博士后科学基金资助项目(2023M731291)。

摘　　要：通过棋盘试验、生长曲线试验、杀菌曲线试验、细菌膜通透性检测试验、外排泵功能检测试验、细胞毒性试验以及大蜡螟存活率试验等筛选出TMexCD1-TOprJ1外排泵抑制剂并验证其体内外替加环素协同抗菌活性。结果显示,白屈菜(Herba chelidonii)中活性成分盐酸白屈菜红碱显著提高替加环素的抗肺炎克雷伯菌(Klebsiella pneumoniae,K.pneumoniae)活性(FICI≤0.5),通过与TMexC1/TMexD1/TOprJ1活性位点结合,抑制细菌外排泵的功能,同时增强替加环素对细菌膜的损伤,从而发挥与替加环素协同杀菌的作用;盐酸白屈菜红碱对J774A.1、Hep-2细胞无毒性作用。结果表明,盐酸白屈菜红碱作为一种新型替加环素增效剂具有抗击高致病性K.pneumoniae感染的潜力,为K.pneumoniae感染治疗提供了候选先导化合物。The aim of this study was to screen the TMexCD1-TOprJ1efflux pump inhibitor and verify its synergistic antimicrobial activity of tigecycline in vivo and in vitro by using checkerboard assay,growth curve assay,time-killing assay,bacterial membrane permeability assay,efflux pump function assay,cytotoxicity assay,and survival rate of Galleria mellonella assay.The active ingredient in Herba chelidonii,chelerythrine chloride,significantly increased the anti-K.pneumoniae activity of tigecycline(FICI≤0.5),inhibited the function of the bacterial efflux pumps by binding to the active sites of TMexC1/TMexD1/TOprJ1,and enhanced the damage of tigecycline to bacterial membranes,thus exerting a synergistic effect with tigecycline.Chelerythrine chloride had no toxic effects on J774A.1and Hep-2cells.These findings reveal the potential of chelerythrine chloride as a novel tigecycline adjuvant against highly pathogenic K.pneumoniae infections and provide a candidate lead compound for the treatment of K.pneumoniae infections.

关 键 词：盐酸白屈菜红碱 替加环素 TMexCD1-TOprJ1 协同活性 肺炎克雷伯菌 

分 类 号：S859[农业科学—临床兽医学]