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合成抗菌肽GH12对口腔链球菌的活性

Activity of Synthetic Antimicrobial Peptide GH12 against Oral Streptococci.

作者信息

Tu Huanxin, Fan Yingying, Lv Xueping, Han Sili, Zhou Xuedong, Zhang Linglin

机构信息

State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, People's Republic of China.

出版信息

Caries Res. 2016;50(1):48-61. doi: 10.1159/000442898. Epub 2016 Feb 10.

Abstract

Controlling the growth of cariogenic microorganisms such as oral streptococci is an adjunct therapy for caries-active individuals to prevent and treat caries. Here we investigated the antimicrobial activity of the synthetic amphipathic α- helical antimicrobial peptide GH12 (GLLWHLLHHLLH-NH2) against oral streptococci in vitro. Circular dichroism studies showed that GH12 takes on an α-helical conformation in the presence of membrane-mimicking solvents, and reversed-phase high-performance liquid chromatography studies showed that GH12 remains stable in saliva. The peptide showed bactericidal activity against oral streptococci, with minimum inhibitory concentrations ranging from 6.7 to 32.0 μg/ml. GH12 concentrations 4-fold higher than the minimum bactericidal concentration completely killed oral streptococci within 20 min. Treating oral streptococci with GH12 caused noticeable changes in bacterial viability and morphology based on confocal laser scanning microscopy and scanning electron microscopy. Effects of GH12 on biofilm formation and on viability of mature biofilm were quantified by crystal violet staining and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. GH12 effectively inhibited biofilm formation and metabolic activity in biofilms of oral streptococci, especially S. mutans, S. sobrinus and S. salivarius. These results suggest that GH12 shows rapid and strong antimicrobial activity against oral streptococci in vitro, opening the door to preclinical and clinical studies to explore its potential for caries prevention and treatment.

摘要

控制致龋微生物(如口腔链球菌)的生长是对龋活跃个体进行预防和治疗龋齿的辅助疗法。在此,我们研究了合成的两亲性α-螺旋抗菌肽GH12(GLLWHLLHHLLH-NH2)对口腔链球菌的体外抗菌活性。圆二色性研究表明,GH12在存在模拟膜的溶剂时呈现α-螺旋构象,反相高效液相色谱研究表明,GH12在唾液中保持稳定。该肽对口腔链球菌显示出杀菌活性,最低抑菌浓度范围为6.7至32.0μg/ml。比最低杀菌浓度高4倍的GH12浓度在20分钟内完全杀死口腔链球菌。基于共聚焦激光扫描显微镜和扫描电子显微镜,用GH12处理口腔链球菌会导致细菌活力和形态发生明显变化。通过结晶紫染色和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐试验对GH12对生物膜形成和成熟生物膜活力的影响进行了定量。GH12有效抑制口腔链球菌生物膜的形成和生物膜中的代谢活性,尤其是变形链球菌、远缘链球菌和唾液链球菌。这些结果表明,GH12在体外对口腔链球菌显示出快速且强大的抗菌活性,为探索其在龋齿预防和治疗方面的潜力的临床前和临床研究打开了大门。

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