Advanced Therapies Group, Cardiff University School of Dentistry, Cardiff, United Kingdom.
School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff, United Kingdom.
Antimicrob Agents Chemother. 2018 Apr 26;62(5). doi: 10.1128/AAC.02318-17. Print 2018 May.
plays a major role in many chronic infections. Its ability to readily form biofilms contributes to its success as an opportunistic pathogen and its resistance/tolerance to antimicrobial/antibiotic therapy. A low-molecular-weight alginate oligomer (OligoG CF-5/20) derived from marine algae has previously been shown to impair motility in biofilms and disrupt pseudomonal biofilm assembly. As these bacterial phenotypes are regulated by quorum sensing (QS), we hypothesized that OligoG CF-5/20 may induce alterations in QS signaling in QS regulation was studied by using CV026 biosensor assays that showed a significant reduction in acyl homoserine lactone (AHL) production following OligoG CF-5/20 treatment (≥2%; < 0.05). This effect was confirmed by liquid chromatography-mass spectrometry analysis of C-AHL and 3-oxo-C-AHL production (≥2%; < 0.05). Moreover, quantitative PCR showed that reduced expression of both the and systems was induced following 24 h of treatment with OligoG CF-5/20 (≥0.2%; < 0.05). Circular dichroism spectroscopy indicated that these alterations were not due to steric interaction between the AHL and OligoG CF-5/20. Confocal laser scanning microscopy (CLSM) and COMSTAT image analysis demonstrated that OligoG CF-5/20-treated biofilms had a dose-dependent decrease in biomass that was associated with inhibition of extracellular DNA synthesis (≥0.5%; < 0.05). These changes correlated with alterations in the extracellular production of the pseudomonal virulence factors pyocyanin, rhamnolipids, elastase, and total protease ( < 0.05). The ability of OligoG CF-5/20 to modify QS signaling in PAO1 may influence critical downstream functions such as virulence factor production and biofilm formation.
在许多慢性感染中发挥着重要作用。它形成生物膜的能力使其成为机会性病原体的成功,并使其具有对抗生素/抗生素治疗的耐药性/耐受性。一种来自海洋藻类的低分子量海藻酸盐低聚物(OligoG CF-5/20)先前已被证明可损害生物膜中的运动性并破坏假单胞菌生物膜组装。由于这些细菌表型受群体感应(QS)调节,我们假设 OligoG CF-5/20 可能会改变 QS 信号在 QS 调节中的作用。通过使用 CV026 生物传感器测定法研究了 QS 调节,该测定法表明 OligoG CF-5/20 处理后酰基高丝氨酸内酯(AHL)的产生显着降低(≥2%; <0.05)。通过 C-AHL 和 3-氧代-C-AHL 产生的液相色谱-质谱分析证实了这种作用(≥2%; <0.05)。此外,定量 PCR 显示,在用 OligoG CF-5/20 处理 24 小时后,两个 和 系统的表达均降低(≥0.2%; <0.05)。圆二色性光谱表明,这些变化不是由于 AHL 和 OligoG CF-5/20 之间的空间相互作用引起的。共焦激光扫描显微镜(CLSM)和 COMSTAT 图像分析表明,OligoG CF-5/20 处理的生物膜的生物量呈剂量依赖性降低,这与抑制细胞外 DNA 合成有关(≥0.5%; <0.05)。这些变化与假单胞菌毒力因子绿脓菌素,鼠李糖脂,弹性蛋白酶和总蛋白酶的细胞外产生的变化相关( <0.05)。OligoG CF-5/20 修饰 PAO1 中 QS 信号的能力可能会影响关键的下游功能,例如毒力因子的产生和生物膜的形成。