亚致死性苯丙酮介导的抗菌光动力疗法后大肠杆菌的应激反应:RNA-Seq 研究。

IF 2.7 3区 化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Photochemical & Photobiological Sciences Pub Date : 2024-08-01 Epub Date: 2024-08-05 DOI:10.1007/s43630-024-00617-3
Denise Muehler, Silvia Morini, Janina Geißert, Christina Engesser, Karl-Anton Hiller, Matthias Widbiller, Tim Maisch, Wolfgang Buchalla, Fabian Cieplik
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引用次数: 0

摘要

由于抗菌光动力疗法(aPDT)的适应性和细菌应激反应背后的分子机制尚不完全清楚,本研究的目的是利用 RNA 测序(RNA-Seq)技术研究大肠杆菌在经 aPDT 亚致死处理后的转录组应激反应。使用亚致死剂量的光敏剂 SAPYR 对静止期大肠杆菌的浮游培养物进行 aPDT 处理。处理后提取 RNA,并在 Illumina NextSeq 500 上进行 RNA-Seq 分析。通过 qRT-PCR 对差异表达基因进行了分析和验证。根据通路富集分析对差异基因表达进行的分析表明,在使用 aPDT 进行亚致死处理后,大肠杆菌中有大量基因和通路被显著上调或下调。与辐照对照组相比,经 aPDT 亚致死处理后,1018 个基因的表达上调,648 个基因的表达下调。对差异表达基因和明显失调途径的分析表明,涉及氧化应激反应和细菌膜损伤的基因受到调控。总之,研究结果表明,在使用 SAPYR 暴露于 aPDT 时,大肠杆菌出现了转录组应激反应,并揭示了可能导致适应性发展的潜在分子机制。
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Stress response in Escherichia coli following sublethal phenalene-1-one mediated antimicrobial photodynamic therapy: an RNA-Seq study.

Since the molecular mechanisms behind adaptation and the bacterial stress response toward antimicrobial photodynamic therapy (aPDT) are not entirely clear yet, the aim of the present study was to investigate the transcriptomic stress response in Escherichia coli after sublethal treatment with aPDT using RNA sequencing (RNA-Seq). Planktonic cultures of stationary phase E. coli were treated with aPDT using a sublethal dose of the photosensitizer SAPYR. After treatment, RNA was extracted, and RNA-Seq was performed on the Illumina NextSeq 500. Differentially expressed genes were analyzed and validated by qRT-PCR. Furthermore, expression of specific stress response proteins was investigated using Western blot analysis.The analysis of the differential gene expression following pathway enrichment analysis revealed a considerable number of genes and pathways significantly up- or down-regulated in E. coli after sublethal treatment with aPDT. Expression of 1018 genes was up-regulated and of 648 genes was down-regulated after sublethal treatment with aPDT as compared to irradiated controls. Analysis of differentially expressed genes and significantly de-regulated pathways showed regulation of genes involved in oxidative stress response and bacterial membrane damage. In conclusion, the results show a transcriptomic stress response in E. coli upon exposure to aPDT using SAPYR and give an insight into potential molecular mechanisms that may result in development of adaptation.

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来源期刊
Photochemical & Photobiological Sciences
Photochemical & Photobiological Sciences 生物-生化与分子生物学
CiteScore
5.60
自引率
6.50%
发文量
201
审稿时长
2.3 months
期刊介绍: A society-owned journal publishing high quality research on all aspects of photochemistry and photobiology.
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