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微生物学通报

抗菌肽17BIPHE2对金黄色葡萄球菌生物被膜的抑制作用
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国家自然科学基金项目(No. 31360622);宁夏临床病原微生物重点实验室开放课题项目(No. LCPM201501)


Inhibition of Staphylococcus aureus biofilm by antimicrobial peptide 17BIPHE2
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    摘要:

    【目的】研究抗菌肽17BIPHE2单独使用及联合抗生素对金黄色葡萄球菌(Staphylococcus aureus)生物被膜的抑制作用。【方法】采用刚果红平板测试法和结晶紫染色评估受试菌形成生物被膜的能力;微量肉汤稀释法和琼脂平板测试法测定金黄色葡萄球菌最小抑菌浓度(MIC)和最小杀菌浓度(MBC);利用抑制金黄色葡萄球菌黏附实验和生物被膜形成抑制实验观察17BIPHE2单独使用及联合抗生素对生物被膜黏附阶段和形成阶段的影响;通过扫描电子显微镜(SEM)观察17BIPHE2单独使用及联合抗生素对成熟生物被膜的清除作用。【结果】17BIPHE2 的MIC为8 μmol/L,1/2×MIC就可以有效抑制浮游菌的生长。单独使用17BIPHE2在细菌黏附阶段抑制率为40%,在生物被膜形成阶段抑制率达到35%。17BIPHE2联合抗生素使用较单独使用抗生素其抑制率均有所下降。生物被膜成熟阶段17BIPHE2于1/4×MIC浓度即可促进生物被膜崩解,1×MIC生物被膜崩解同时细菌黏附量有所下降,联合万古霉素促进生物被膜崩解同时细菌胞质大量外泄。【结论】抗菌肽17BIPHE2具有良好的抑制金黄色葡萄球菌生物被膜作用,联合抗生素其抗生物被膜作用进一步提高。这将为治疗由金黄色葡萄球菌生物被膜引起的相关感染提供了一个新思路。

    Abstract:

    [Objective] To study the inhibition of staphylococcus aureus (S. aureus) biofilm by antimicrobial peptide 17BIPHE2 used alone or combined with antibiotics. [Methods] Congo red plate test and crystal violet staining test were used to assess the ability of biofilm formation by S. aureus. Minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of S. aureus were tested by broth microdilution method and agar plate method. The bacteria adhension was observed by S. aureus adhesion test and biofilm formation was analyzed by biofilm formation test. They were treated by 17BIPHE2 alone or combined with antibiotics. The removal of mature biofilm was observed by scanning electron microscope (SEM) after the treatment of 17BIPHE2 alone or combined with antibiotics. [Results] When MIC of 17BIPHE2 was 8 μmol/L, 1/2×MIC could inhibit the growth of planktonic bacteria effectively. In the adhension stage, the inhibition rate was 40% by 17BIPHE2 treated only. In the biofilm formation stage, the inhibition rate was 35%. Further, the rates for both stages decreased if combined with antibiotics. Antimicrobial peptide 17BIPHE2 could promote biofilm disintegration in the mature stage at 1/4×MIC concentration. Biofilm disintegration and bacterial adhesion could decrease at 1×MIC concentration. Furthermore, the combination of vancomycin promoted disintegration of bacterial biofilm as well as lots of leakage. [Conclusion] Antimicrobial peptides 17BIPHE2 was able to inhibit biofilm effectively, especially combined with antibiotics, could promote the inhibition more effectively. This article provides a new idea for treatment of S. aureus biofilm related infection.

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回丽媛,李军,朱明星,王秀青. 抗菌肽17BIPHE2对金黄色葡萄球菌生物被膜的抑制作用[J]. 微生物学通报, 2017, 44(4): 872-880

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  • 在线发布日期: 2017-04-05
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