Huo Lijun, Huang Xiangya, Ling Junqi, Liu Hongyan, Liu Jia
Department of Operative Dentistry, Preventive Dentistry and Endodontics, The Affiliated Stomatology Hospital of Kunming Medical University, Kunming, Yunnan 650500, P.R. China.
Department of Operative Dentistry, Preventive Dentistry and Endodontics, Affiliated Stomatology Hospital of Sun Yat-sen University, Guangzhou, Guandong 510055, P.R. China.
Exp Ther Med. 2018 Feb;15(2):1886-1893. doi: 10.3892/etm.2017.5631. Epub 2017 Dec 14.
The present study aimed to design, synthesize and screen specifically targeted antimicrobial peptides (STAMPs) that can selectively kill () in the biofilm, and to detect protein metabolism, in order to investigate the mechanism of the antibacterial functions of STAMPs against . A series of STAMPs were synthesized, and their effects on the selective antibacterial activity of on single species and multi-species biofilms under the condition of the planktonic state were studied. The total protein of was extracted before and after C11H, and matrix-assisted laser adsorption ionization-time of flight mass spectrometry identification was performed. The antibacterial activity on planktonic was increased 3- to 4-fold via C8H, C11H, C12H, C13H, and C14H compared with hLF1-11 (H) alone, and there was no difference between () and (). C8H, C11H, C12H, C13H, and C14H had significant inhibitory effects on the growth of biofilm, but there were no significant effects on and biofilms. The number of in biofilm decreased at 4 h after C8H, C11H, C12H, C13H and C14H and C8, C11, C12, C13 and C14 had no effect on the growth of planktonic and biofilm states of , and species. C11H and C12H exhibited the most obvious effects, followed by C13H and C14H, and then C8H. A total of 21 protein spots with a mean change ratio of 1.5 were identified, all of which were downregulated after C11H. A total of 19 proteins were successfully identified, including cell cycle-relative proteins, nucleic acid metabolism-related enzymes and proteins, virulence factors, protein biosynthesis and regulation, proteins involved in energy metabolism, and proteins with unknown function. In the present study, STAMPs with selective antibacterial activity against grown in planktonic or biofilm states but without obvious effects on oral and multi-species biofilm were successfully designed and synthesized. Differential protein expression before and after C11H was identified. The mechanism of the antibacterial function was also discussed. Results of the present study laid the foundation for application of STAMPs in the prevention and treatment of dental caries.
本研究旨在设计、合成并筛选能够选择性杀死生物膜中（）的特异性靶向抗菌肽（STAMPs），并检测蛋白质代谢，以探究STAMPs对（）抗菌功能的作用机制。合成了一系列STAMPs，并研究了它们在浮游状态下对单一物种和多物种生物膜中（）的选择性抗菌活性的影响。在C11H处理前后提取（）的总蛋白，并进行基质辅助激光吸附电离飞行时间质谱鉴定。与单独的hLF1-11（H）相比，C8H、C11H、C12H、C13H和C14H使浮游（）的抗菌活性提高了3至4倍，且（）与（）之间无差异。C8H、C11H、C12H、C13H和C14H对（）生物膜的生长具有显著抑制作用，但对（）和（）生物膜无显著影响。C8H、C11H、C12H、C13H和C14H处理4小时后，生物膜中（）的数量减少，且C8、C11、C12、C13和C14对（）、（）和（）物种的浮游和生物膜状态的生长无影响。C11H和C12H表现出最明显的效果，其次是C13H和C14H，然后是C8H。共鉴定出21个平均变化率为1.5的蛋白斑点，所有这些斑点在C11H处理后均下调。成功鉴定出19种蛋白质，包括细胞周期相关蛋白、核酸代谢相关酶和蛋白质、毒力因子、蛋白质生物合成和调控、能量代谢相关蛋白质以及功能未知的蛋白质。在本研究中，成功设计并合成了对浮游或生物膜状态下生长的（）具有选择性抗菌活性但对口腔（）和多物种生物膜无明显影响的STAMPs。鉴定了C11H处理前后的差异蛋白表达。还讨论了抗菌功能的机制。本研究结果为STAMPs在龋齿防治中的应用奠定了基础。
