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人源抗菌肽LL-37增强表没食子儿茶素没食子酸酯对变形链球菌的抗生物膜作用。

Human cathelicidin LL-37 enhance the antibiofilm effect of EGCG on Streptococcus mutans.

作者信息

Guo Yi-Jie, Zhang Bo, Feng Xue-Song, Ren Hui-Xun, Xu Ji-Ru

机构信息

Department of Pathogenic Microbiology and Immunology, School of Basic Medical Sciences, Xi'an Jiaotong University, Yanta West Road No.76, Xi'an, 710061, ShaanXi, China.

Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education of China, Xi'an Jiaotong University, Yanta West Road No.76, Xi'an, 710061, ShaanXi, China.

出版信息

BMC Oral Health. 2016 Sep 22;16(1):101. doi: 10.1186/s12903-016-0292-y.

Abstract

BACKGROUND

Streptococcus mutans forms biofilms as a resistance mechanism against antimicrobial agents in the human oral cavity. We recently showed that human cathelicidin LL-37 exhibits inhibitory effects on biofilm formation of S. mutans through interaction with lipoteichoic acid (LTA), but without antibacterial or biofilm dispersal abilities. (-)-Epigallocatechin gallate (EGCG) is the most abundant constituent of tea catechins that has the greatest anti-infective potential to inhibit the growth of various microorganisms and biofilm formation. Therefore, in this study, we evaluated whether LL-37 interacts with EGCG to enhance the antibiofilm effect of EGCG on S. mutans biofilm formation.

METHODS

Clinical S. mutans strains (n = 10) isolated from children's saliva were tested in a biofilm formation assay. The antibiofilm effect of EGCG with and without LL-37 was analyzed by the minimum biofilm eradication concentration assay and confirmed using field emission-scanning electron microscopy. In addition, the interaction among EGCG, LL-37, and LTA of S. mutans was determined using quartz crystal microbalance analysis.

RESULTS

EGCG killed 100 % of planktonic S. mutans within 5 h, inhibited biofilm formation within 24 h, and reduced bacteria cells in preformed biofilms within 3 h at a concentration of 0.2 mg/mL. However, EGCG did not appear to interact with LTA. LL-37 effectively enhanced the bactericidal activity of EGCG against biofilm formation and preformed biofilms as determined by quantitative crystal violet staining and field emission-scanning electron microscopy. In addition, quartz crystal microbalance analysis revealed that LL-37 interacted with EGCG and promoted binding between EGCG and LTA of S. mutans.

CONCLUSIONS

We show that LL-37 enhances the antibiofilm effect of EGCG on S. mutans. This finding provides new knowledge for dental treatment by using LL-37 as a potential antibiofilm compound.

摘要

背景

变形链球菌形成生物膜作为其在人类口腔中对抗抗菌剂的一种抗性机制。我们最近发现,人源抗菌肽LL-37通过与脂磷壁酸(LTA)相互作用,对变形链球菌的生物膜形成具有抑制作用,但不具备抗菌或生物膜分散能力。(-)-表没食子儿茶素没食子酸酯(EGCG)是茶儿茶素中含量最丰富的成分,具有抑制多种微生物生长和生物膜形成的最大抗感染潜力。因此,在本研究中,我们评估了LL-37是否与EGCG相互作用以增强EGCG对变形链球菌生物膜形成的抗生物膜作用。

方法

从儿童唾液中分离出的10株临床变形链球菌菌株用于生物膜形成试验。通过最小生物膜清除浓度试验分析了有无LL-37时EGCG的抗生物膜作用,并使用场发射扫描电子显微镜进行了确认。此外,利用石英晶体微天平分析确定了EGCG、LL-37和变形链球菌LTA之间的相互作用。

结果

EGCG在5小时内杀死了100%的浮游变形链球菌,在24小时内抑制生物膜形成,并在3小时内以0.2mg/mL的浓度减少了预先形成的生物膜中的细菌细胞。然而,EGCG似乎不与LTA相互作用。通过定量结晶紫染色和场发射扫描电子显微镜测定,LL-37有效地增强了EGCG对生物膜形成和预先形成的生物膜的杀菌活性。此外,石英晶体微天平分析表明,LL-37与EGCG相互作用,并促进了EGCG与变形链球菌LTA之间的结合。

结论

我们发现LL-37增强了EGCG对变形链球菌的抗生物膜作用。这一发现为将LL-37用作潜在的抗生物膜化合物进行牙科治疗提供了新知识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9349/5034579/9135863d395c/12903_2016_292_Fig1_HTML.jpg

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