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亚致死浓度苯并[a]蒽酮介导的光动力抗菌治疗后大肠杆菌的应激反应:一项 RNA-Seq 研究。

Stress response in Escherichia coli following sublethal phenalene-1-one mediated antimicrobial photodynamic therapy: an RNA-Seq study.

机构信息

Department of Conservative Dentistry and Periodontology, University Hospital Regensburg, Regensburg, Germany.

Quantitative Biology Center (QBiC), University of Tübingen, Tübingen, Germany.

出版信息

Photochem Photobiol Sci. 2024 Aug;23(8):1573-1586. doi: 10.1007/s43630-024-00617-3. Epub 2024 Aug 5.

Abstract

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.

摘要

由于适应的分子机制以及细菌对抗生素光动力疗法(aPDT)的应激反应的机制尚不完全清楚,本研究的目的是使用 RNA 测序(RNA-Seq)研究亚致死剂量的 PDT 处理后大肠杆菌中的转录组应激反应。使用亚致死剂量的光敏剂 SAPYR 对处于静止期的大肠杆菌浮游培养物进行 PDT 处理。处理后,提取 RNA,并在 Illumina NextSeq 500 上进行 RNA-Seq。通过 qRT-PCR 分析和验证差异表达基因。此外,使用 Western blot 分析研究了特定应激反应蛋白的表达。

对差异表达基因进行通路富集分析的结果表明,亚致死剂量的 PDT 处理后,大肠杆菌中有相当数量的基因和通路显著上调或下调。与辐照对照相比,亚致死剂量的 PDT 处理后 1018 个基因上调,648 个基因下调。差异表达基因和显著失调通路的分析表明,与氧化应激反应和细菌膜损伤相关的基因受到调控。

总之,这些结果表明,SAPYR 用于 aPDT 会导致大肠杆菌产生转录组应激反应,并深入了解可能导致适应发展的潜在分子机制。

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