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首页> 美国卫生研究院文献>other >Transcriptional Modulation of Penicillin-Binding Protein 1b Outer Membrane Protein P2 and Efflux Pump (AcrAB-TolC) during Heat Stress Is Correlated to Enhanced Bactericidal Action of Imipenem on Non-typeable Haemophilus influenzae
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Transcriptional Modulation of Penicillin-Binding Protein 1b Outer Membrane Protein P2 and Efflux Pump (AcrAB-TolC) during Heat Stress Is Correlated to Enhanced Bactericidal Action of Imipenem on Non-typeable Haemophilus influenzae

机译：在热应激过程中青霉素结合蛋白1b外膜蛋白P2和外排泵（AcrAB-TolC）的转录调节与亚胺培南对非典型流感嗜血杆菌的杀菌作用增强相关。

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>Objective: The purpose of the present study was to investigate the penicillin binding proteins (PBPs), drug influx and efflux modulations during heat stress and their effects on the bactericidal action of imipenem on non-typeable Haemophilus influenzae (NTHi).>Methods: The two NTHi clinical isolates (GE47 and GE88, imipenem MICs by E-test > 32 μg/mL) examined in this study were collected at Geneva University Hospitals. The imipenem killing activity was assessed after incubation of the NTHi strains at either 37 or 42°C for 3 h with increasing concentrations of imipenem. The detection of PBPs was carried out by Bocillin-FL. Global transcriptional changes were monitored by RNA-seq after pre-incubation of bacterial cells at either 37 or 42°C, and the expression levels of relevant target genes were confirmed by qRT-PCR.>Results: Quantitation of NTHi viable cells after incubation with 0.25 μg/mL of imipenem for 3 h revealed more than a twofold decrease in GE47 and GE88 viable cells at 42°C as compared to 37°C. Transcriptome analysis showed that under heat stress conditions, there were 141 differentially expressed genes with a | log2(fold change)| > 1, including 67 up-regulated and 74 down-regulated genes. The expression levels of ponB (encoding PBP1b) and acrR (regulator of AcrAB-TolC efflux pump) were significantly increased at 42°C. In contrast, the transcript levels of ompP2 (encoding the outer membrane protein P2) and acrB gene (encoding AcrB) were significantly lower under heat stress condition.>Conclusion: This study shows that the transcriptional modulation of ponB, ompP2, acrR, and acrB in the heat stress response is correlated to enhanced antimicrobial effects of imipenem on non-typeable H. influenzae.
机译：>目的：本研究的目的是研究热应激过程中的青霉素结合蛋白（PBP），药物流入和流出调节及其对亚胺培南对不可分型流感嗜血杆菌的杀菌作用的影响（ NTHi）。>方法：本研究中检测的两种NTHi临床分离株（GE47和GE88，亚胺培南MIC，通过E-test> 32μg/ mL）均收集在日内瓦大学医院。在NTHi菌株于37或42°C孵育3小时后，随着亚胺培南浓度的增加，评估亚胺培南的杀伤活性。 PBP的检测由Bocillin-FL进行。在37或42°C的温度下预孵育细菌细胞后，通过RNA-seq监测全局转录变化，并通过qRT-PCR确认相关靶基因的表达水平。>结果：与0.25μg/ mL亚胺培南孵育3小时后，NTHi活细胞在42°C下的GE47和GE88活细胞比37°C减少了两倍以上。转录组分析显示，在热胁迫条件下，存在141个差异表达基因，其中| log2（倍数变化）| > 1，包括67个上调基因和74个下调基因。在42°C，ponB（编码PBP1b）和acrR（AcrAB-TolC外排泵的调节剂）的表达水平显着增加。相比之下，在热应激条件下，ompP2（编码外膜蛋白P2）和acrB基因（编码AcrB）的转录水平显着降低。>结论：该研究表明，ponB的转录调控， ompP2，acrR和acrB在热应激反应中与亚胺培南对不可分型流感嗜血杆菌的增强抗菌作用相关。

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