In vitro activity of tigecycline and proteomic analysis of tigecycline adaptation strategies in clinical Enterococcus faecalis isolates from China.
In vitro activity of tigecycline and proteomic analysis of tigecycline adaptation strategies in clinical Enterococcus faecalis isolates from China.
复制标题
中国临床粪肠球菌替加环素的体外活性和替加环素适应策略的蛋白质组学分析。
DOI：
10.1016/j.jgar.2022.04.022
复制
发表时间：
2022
影响因子：
4.6
通讯作者：
Wen,Zewen
中科院分区：
文献类型：
--
作者：
Bai,Bing;Chen,Chengchun;Zhao,Yuxi;Xu,Guangjian;Yu,Zhijian;Tam,VincentH;Wen,Zewen
目的探讨替加环素（TGC）在粪肠球菌（enterococcus faecalisischina）体内的体外活性及TGC胁迫反应和耐药性的分子机制。方法TGC对399E菌的抗生素敏感性和抗菌活性。对分离粪杆菌进行评价。用群体分析谱法评价杂种抗性。通过鉴定四环素（tet）靶点的基因突变和外排蛋白抑制剂（EPIs）的分析，研究了耐药和异源耐药机制。此外，定量蛋白质组学研究了e的全球蛋白质组学反应。TGC体外诱导抗性分离株的耐药机制。结果stgc对临床ale的最低抑菌浓度（mic）。粪杆菌≤0.5 mg/L。TGC对生物膜的形成有明显的抑制作用。TGC异抗发生率为1.75% (7/399)，EPI可逆转异抗衍生无性系TGC MIC值的升高。TGC耐药与16S rRNA位点或30S核糖体蛋白S10突变有关。暴露于1/2倍MIC浓度的TGC后，分别鉴定出105个和356个差异表达蛋白，而TGC耐药分离物中鉴定出356个差异表达蛋白。差异表达蛋白在翻译和DNA复制过程中富集。此外，多种三磷酸腺苷（ATP）结合盒（ABC）转运蛋白上调。结论stgc对中国大部分临床分离株具有良好的抑菌活性。然而,E。粪便在TGC暴露过程中表现出较强的适应机制：TGC选择导致TGC靶位点突变，外排泵表达升高，从而产生TGC抗性。
ObjectivesThis study aimed to investigate the in vitro activities of tigecycline (TGC) and the underlying molecular mechanisms of TGC stress response and resistance in clinicalEnterococcus faecalisisolates from China.MethodsAntimicrobial susceptibility and antibiofilm activities of TGC in 399E. faecalisisolates were evaluated. Heteroresistance was evaluated by population analysis profiling. Resistance and heteroresistance mechanisms were investigated by identifying genetic mutations in tetracycline (tet) target sites and through analysis of efflux protein inhibitors (EPIs). Furthermore, quantitative proteomics was used to investigate the global proteomic response ofE. faecalisto TGC stress, as well as the resistance mechanisms of TGC within in vitro induced resistant isolate.ResultsTGC minimum inhibitory concentrations (MICs) against clinicalE. faecalisisolates were ≤0.5 mg/L. TGC displayed remarkable inhibitory activity against biofilm formation. The occurrence rate of TGC heteroresistance was 1.75% (7/399), and the increased TGC MIC values of heteroresistance-derived clones could be reversed by EPI. TGC resistance was associated with mutations in the 16S rRNA site or 30S ribosomal protein S10. A total of 105 and 356 differentially expressed proteins was identified after being exposed to 1/2× MIC concentrations of TGC, while 356 differentially expressed proteins was identified in TGC-resistant isolate. The differentially expressed proteins were enriched in the translation and DNA replication process. In addition, multiple adenosine triphosphate (ATP)-binding cassette (ABC) transporters were upregulated.ConclusionsTGC exhibited excellent activity against a substantial proportion of clinical isolates from China. However,E. faecalisexhibited a strong adaptation mechanism during TGC exposure: mutation of TGC target sites and elevated expression of efflux pumps under TGC selection, resulting in TGC resistance.