抗菌肽/三氯生共修饰的金纳米颗粒用于增强抗菌效果的研究  

Antibacterial peptide/triclosan dual functionalized gold nanoparticles for enhanced antibacterial effect

作　　者：王婧[1] 蔡晓瑤 杨丽军[1] 黄帆[1] 刘鉴峰[1] WANG Jing;CAI Xiao-yao;YANG Li-jun;HUANG Fan;LIU Jian-feng(Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin 300192, China;The Sixth Mobile Detachment of the First Mobile Command of the Armed Police Force, Baoding 072450, Hebei, China)

机构地区：[1]北京协和医学院.中国医学科学院放射医学研究所天津市放射医学与分子核医学重点实验室,天津300192 [2]武警第一机动总队机动第六支队,河北保定072450

出　　处：《生物医学工程与临床》2019年第3期252-257,共6页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(21708048);中国医学科学院医学与健康科技创新工程项目(2016-I2M-3-022;2017-I2M-2-004);中央高校基本科研业务费专项资金资助(3332018194;3332018115)

摘　　要：目的设计合成一类新型抗菌肽/三氯生双修饰金纳米颗粒(Au-TCS-RWNPs)抗菌剂,对其纳米结构进行表征,并研究其体外抗菌活性和血液相容性。方法先利用柠檬酸盐还原法合成金纳米颗粒(AuNPsh通过将抗菌肽精氨酸-色氨酸-精氨酸-色氨酸-精氨酸-色氨酸-半胱氨酸(RWRWRWC)和三氣生-甘氨酸-甘氨酸-半胱氨酸(TCS-GGC)共修饰或单独修饰到Au NPs上,得到Au-TCS-RW NPs、Au-TCS NPs、Au-RW NPs。利用动态光散射(DLS)粒度仪和透射电子显微镜<TEM)测定4种Au NPs的粒径和形貌;通过紫外-可见吸收光谱表征4种Au NPs的光学性质;采用电位仪测定4种AuNPs的表面电位;通过生物发光成像系统研究Au-TCS-RWNPs抗生物发光转基因的金黄色葡萄球菌的效果;通过溶血实验表征Au-TCS-RW NPs的血液相容性。结果 Au NPs、Au-TCS NPs、Au-RW NPs、An-TCS-RW NPs分别为直径27.79 nm、28.06 nm、26.19 nm、26.38 rmi的球形纳米粒,表面电荷分布存在明显差异,分别是-2.2 mV、-4.0mV、3.0 mV、4.0mV。Au-TCS-RWNPs对金黄色葡萄球菌的杀伤效果具有浓度依赖性,其抗菌效果优于未修饰的Au NPs、Au-RW NPs及Au-TCS NPs。Au NPs、Au-TCS NPs、Au-RW NPs及Au-TCS-RW NPs在质量浓度范围内(0～400 pg/mL)对红细胞的溶血率均在5%以下。结论够通过修饰的抗菌肽和三氯生发挥协同抗菌效果,并且具有较好的血液相容性,是一种具有临床应用前景的有效纳米抗菌剂。Objective To design and synthesize a new type of antibacterial agent based on antibacterial peptide/triclosan dual functionalized gold nanoparticles(Au-TCS-RW NPs), and characterize its nanostructure, investigate the antibacterial effect and blood compatibility in vitro. Methods The gold nanoparticles(Au NPs) were synthesized by citrate reduction. Au-TCS-RW NPs,Au-TCS NPs and Au-RW NPs were obtained by co-modifying or individually modifying antibacterial peptide arginine-tryptophan-arginine-tryptophan-arginine-tryptophan-cysteine(RWRWRWC) and triclosan-glycine-glycine-cysteine(TCS-GGC) onto Au NPs. The size and morphology of the four kinds of Au NPs were determined by dynamic light scattering(DLS) and transmission electron microscopy(TEM). The optical properties of the four kinds of Au NPs were characterized by UV-visible absorption spectroscopy. Surface potentials of the four kinds of Au NPs was measured by potentiometer. The antibacterial effect of AuTCS-RW NPs on bioluminescent transgenic Staphylococcus aureus was studied by bioluminescent imaging system. The blood compatibility of Au-TCS-RW NPs was characterized by hemolysis test. Results The Au NPs, Au-TCS NPs, Au-RW NPs and Au-TCS-RW NPs were spherical nanoparticles with diameters of 27.79 nm, 28.06 nm, 26.19 nm and 26.38 nm, respectively, and the surface charge distributions were significantly different, which were-2.2 mV,-4.0 mV, 3.0 mV and 4.0 mV, respectively. The antibacterial effect of Au-TCS-RW NPs on Staphylococcus aureus was concentration dependent, and the effect was superior to that of unmodified Au NPs, Au-RW NPs modified with antibacterial peptides alone and Au-TCS NPs modified with triclosan alone. The hemolysis rates of Au NPs, Au-TCS NPs, Au-RW NPs and Au-TCS-RW NPs in concentration range(0-400 μg/mL) were all less than 5 %. Conclusion It is demonstrated that Au-TCS-RW NPs could exert synergistic antibacterial effect through modified antibacterial peptide/triclosan, and shows good blood compatibility, which act as an efficient nano antibact

关 键 词：抗菌肽 三氯生 金纳米颗粒 联合治疗 抗菌 

分 类 号：R318.08[医药卫生—生物医学工程]