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香芹酚抑制金黄色葡萄球菌生物被膜的形成
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国家自然科学基金(31702284);四川农业大学双支计划(03571444);国家农业产业技术体系四川兽药创新团队专项(CARS-SVDIP)


Inhibition of Staphylococcus aureus biofilm by carvacrol
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    摘要:

    【背景】生物被膜是细菌的一种自我保护形式,可以增强细菌对药物及宿主免疫应答的抵抗力,引起细菌耐药性和持续性感染。【目的】探究香芹酚对金黄色葡萄球菌生物被膜的作用机制,为开发新型抗生物被膜药物提供可靠的理论依据。【方法】通过结晶紫染色法检测香芹酚对供试菌株生物被膜形成的抑制和对成熟生物被膜的清除作用;使用刚果红平板法探究香芹酚对供试菌株生物被膜形成过程中细胞间多糖黏附素(polysaccharide intercellular adhesion,PIA)合成的作用;通过分光光度法检测香芹酚对胞外DNA (extracellular DNA,eDNA)分泌的抑制作用;利用RT-PCR技术检测香芹酚对供试菌株的生物被膜相关基因icaA、cidA和sarA转录水平的影响。【结果】香芹酚对生物被膜形成的抑制和生物被膜的清除均有较强作用效果。256 μg/mL香芹酚抑制PIA合成和eDNA释放的效果显著。香芹酚可通过抑制相关基因转录从而抑制生物被膜的形成,当64 μg/mL的香芹酚作用后,sarA的转录水平降低了60.44%±2.91%,cidA的转录水平降低了76.48%±1.67%,icaA的转录水平降低了70.00%±1.94%。【结论】香芹酚对金黄色葡萄球菌ATCC 25923生物被膜的抑制和清除作用显著,其作用机制主要是通过降低icaA、sarA和cidA基因转录水平抑制PIA的合成和eDNA的释放。

    Abstract:

    [Background] Biofilm is a form of bacterial self-protection, which can enhance the resistance of bacteria to antibiotics and host immune response, and causes bacterial resistance and persistent infection. [Objective] To provide a reliable theoretical basis for the development of new natural antibacterial drugs, the inhibition mechanism of carvacrol on the biofilm formation of Staphylococcus aureus was explored. [Methods] Crystal violet staining was used to detect the inhibitory effect of carvacrol on biofilm formation and the clear effect on mature biofilm of tested strains. We used congo red agar to detect the influence of carvacrol on polysaccharide intercellular adhesion (PIA) formation, and used spectrophotometer to detect the influence of carvacrol on extracellular DNA (eDNA) release. RT-PCR analysis was used to determine the effect of carvacrol on the transcription levels of icaA, cidA and sarA. [Results] Carvacrol has strong effects on the inhibition of biofilm formation and the removal of mature biofilm. The inhibition of PIA synthesis and eDNA release by carvacrol at 256 μg/mL was significant. Carvacrol can inhibit the formation of biofilm by inhibiting the transcription of related genes. When carvacrol was 64 μg/mL, the transcription level of sarA decreased by 60.44%±2.91%, cidA by 76.48%±1.67%, and icaA decreased by 70.00%±1.94%. [Conclusion] Carvacrol had significant inhibitory and removal effects on the biofilm of Staphylococcus aureus 25923. Its mechanism is to inhibit the synthesis of PIA and the release of eDNA by reducing the transcription levels of icaA, sarA and cidA genes.

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戴雨芸,李超,袁中伟,李欣越,何泾正,尹立子. 香芹酚抑制金黄色葡萄球菌生物被膜的形成[J]. 微生物学通报, 2020, 47(3): 813-820

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  • 在线发布日期: 2020-03-04
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