Effect of the efflux inhibitors 1-(1-naphthylmethyl)-piperazine and phenyl-arginine-β-naphthylamide on antimicrobial susceptibility and virulence factor production in Vibrio cholerae
Effect of the efflux inhibitors 1-(1-naphthylmethyl)-piperazine and phenyl-arginine-β-naphthylamide on antimicrobial susceptibility and virulence factor production in Vibrio cholerae
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DOI：
10.1093/jac/dkn466
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发表时间：
2009-01-01
影响因子：
5.2
通讯作者：
Bina, James E.
中科院分区：
文献类型：
--
作者：
Bina, Xiaowen R.;Philippart, Julie A.;Bina, James E.
本研究的目的是验证外排泵抑制剂（EPIs） 1-（1-萘甲基）-哌嗪（NMP）和苯基精氨酸- β -萘酰胺（PA β N）可以抑制霍乱弧菌耐药-结核分裂（RND）家族外排系统，从而使霍乱弧菌对抗菌药物敏感，并抑制毒力因子霍乱毒素（CT）和毒素协同调节菌毛（TCP）的产生。在NMP和PA β N存在或不存在的情况下，测定了霍乱弧菌对抗菌化合物的敏感性。转录报告器用于评估NMP和PA β N对毒力因子调控因子TcpP和ToxT编码基因表达的影响，而CT和TCP的产生分别使用GM1神经节苷包被微滴板和TcpA Western免疫印迹法通过ELISA检测。NMP和PA β N增强抗菌化合物是霍乱弧菌RND外排系统的底物。PA β N对Triton X-100和脱氧胆酸的RND外排系统有完全抑制作用，但对胆酸和红霉素的外排系统有部分抑制作用。NMP对除SDS外的所有化合物均有部分抑制作用。NMP的存在使SDS的MIC降低到低于RND外排缺陷菌株的水平，而SDS的MIC不受PA β n的影响。EPI没有增强多粘菌素B、青霉素、氨苄西林或氯霉素。NMP和PA β N都抑制了CT和TCP的产生，并且似乎具有独立于RND外排抑制的额外毒力基因抑制活性。RND外排抑制剂代表了霍乱治疗的潜在新疗法。
The aim of the study was to test the hypothesis that the efflux pump inhibitors (EPIs) 1-(1-naphthylmethyl)-piperazine (NMP) and phenyl-arginine-beta-naphthylamide (PA beta N) can inhibit the Vibrio cholerae resistance-nodulation-division (RND) family efflux systems, and thereby render V. cholerae susceptible to antimicrobial agents and inhibit the production of the virulence factors cholera toxin (CT) and the toxin coregulated pilus (TCP).The susceptibility of V. cholerae to antimicrobial compounds was determined in the presence or absence of NMP and PA beta N. Transcriptional reporters were used to assess the effects of NMP and PA beta N on the expression of the genes encoding the virulence factor regulators TcpP and ToxT, whereas CT and TCP production were determined by ELISA using GM1 ganglioside-coated microtitre plates and TcpA Western immunoblotting, respectively.NMP and PA beta N potentiated antimicrobial compounds that were substrates for the V. cholerae RND efflux systems. PA beta N exhibited complete inhibition of the RND efflux systems for Triton X-100 and deoxycholate, but partial inhibition of the efflux systems for cholate and erythromycin. NMP exhibited partial inhibition for all compounds tested except for SDS. The presence of NMP reduced the MIC of SDS to a level that was lower than that observed in an RND efflux-deficient strain, whereas the SDS MIC was unaffected by the presence of PA beta N. Neither EPI potentiated polymyxin B, penicillin, ampicillin or chloramphenicol. Both NMP and PA beta N inhibited the production of CT and the TCP and appeared to have additional virulence gene repressing activity independent of RND efflux inhibition.RND efflux inhibitors represent potential novel therapeutics for the treatment of cholera.