Takayama Saori, Hashimoto Kenji, Kokubu Eitoyo, Taniguchi Masayuki, Tajima Kouji, Ochiai Akihito, Saitoh Eiichi, Saito Atsushi, Ishihara Kazuyuki, Kato Tetsuo
Department of Periodontology, Tokyo Dental College, Tokyo 101-0061, Japan.
Department of Materials Science and Technology, Graduate School of Science and Technology, Niigata University, Niigata 950-2181, Japan.
Microb Pathog. 2016 May;94:2-11. doi: 10.1016/j.micpath.2015.08.011. Epub 2015 Aug 28.
CL(14-25), a dodecapeptide of cyanate lyase from rice, is a novel cationic α-helical antimicrobial peptide. In this study, we examined inhibitory ability of CL(14-25) against endotoxic activities of lipopolysaccharides (LPSs) from Escherichia coli and periodontal pathogenic Aggregatibacter actinomycetemcomitans.
Endotoxin-neutralizing activity of CL(14-25) was evaluated by inhibition to induction of cytokine and nitric oxide in human aortic endothelial cells (HAECs) and RAW264 mouse macrophage cells, respectively. Protective effect of CL(14-25) was determined in mice against lethal toxicity of LPS.
IL-6 in HAECs was induced by stimulation with LPS preparations of A. actinomycetemcomitans and E. coli tested in this study, and addition of CL(14-25) to the medium caused inhibition of their induction in a dose-dependent manner. CL(14-25) inhibited NO induction in RAW264 cells by a smooth type LPS of E. coli O55:B5 and an Rc type LPS of E. coli J5 as well as lipid A of E. coli R515 in a dose-dependent manner. Simultaneous injection of E. coli O55:B5 LPS and CL(14-25) in BALB/c mice resulted in prevention of lethal toxicity of the former. The results of a Limulus amebocyte lysate assay and surface plasmon resonance analysis of interaction between CL(14-25) and E. coli LPS or lipid A showed that CL(14-25) specifically binds to a lipid A moiety of LPS.
The results of present study suggest that CL(14-25) has a potential to be used as a nutraceutical agent for periodontal therapy.
CL(14 - 25)是一种来自水稻氰酸裂解酶的十二肽，是一种新型阳离子α-螺旋抗菌肽。在本研究中，我们检测了CL(14 - 25)对大肠杆菌和牙周病原菌伴放线聚集杆菌脂多糖（LPS）内毒素活性的抑制能力。
分别通过抑制人主动脉内皮细胞（HAECs）中细胞因子的诱导和RAW264小鼠巨噬细胞中一氧化氮的产生，评估CL(14 - 25)的内毒素中和活性。在小鼠中测定CL(14 - 25)对LPS致死毒性的保护作用。
本研究中测试的伴放线聚集杆菌和大肠杆菌的LPS制剂刺激可诱导HAECs中IL-6的产生，向培养基中添加CL(14 - 25)可呈剂量依赖性抑制其诱导。CL(14 - 25)可呈剂量依赖性抑制大肠杆菌O55:B5的光滑型LPS、大肠杆菌J5的Rc型LPS以及大肠杆菌R515的脂质A诱导RAW264细胞产生NO。在BALB/c小鼠中同时注射大肠杆菌O55:B5 LPS和CL(14 - 25)可预防前者的致死毒性。鲎试剂检测以及CL(14 - 25)与大肠杆菌LPS或脂质A相互作用的表面等离子体共振分析结果表明，CL(14 - 25)特异性结合LPS的脂质A部分。
本研究结果表明CL(14 - 25)有潜力用作牙周治疗的营养制剂。
