一种源自水稻氰酸裂解酶的新型阳离子十二肽[CL(14-25)]对大肠杆菌和牙周病原菌伴放线聚集杆菌的脂多糖内毒素活性的抑制作用。

Inhibitory effects of a novel cationic dodecapeptide [CL(14-25)] derived from cyanate lyase of rice on endotoxic activities of LPSs from Escherichia coli and periodontopathic Aggregatibacter actinomycetemcomitans.

作者信息

Takayama Saori, Hashimoto Kenji, Kokubu Eitoyo, Taniguchi Masayuki, Tajima Kouji, Ochiai Akihito, Saitoh Eiichi, Saito Atsushi, Ishihara Kazuyuki, Kato Tetsuo

机构信息

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.

出版信息

Microb Pathog. 2016 May;94:2-11. doi: 10.1016/j.micpath.2015.08.011. Epub 2015 Aug 28.

DOI:10.1016/j.micpath.2015.08.011

PMID:26318142

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

The results of present study suggest that CL(14-25) has a potential to be used as a nutraceutical agent for periodontal therapy.

摘要

目的

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。

方法

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。

结果

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。