Synergistic bacteriostatic properties of 4-hydroxycoumarin-Ag composite
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摘要: 抗生素的滥用导致大量耐药菌出现,对卫生健康和社会经济造成严重威胁,因此,迫切需要开发新型抗菌剂来解决细菌耐药问题。本文以三氟醋酸银(CF3COOAg)为原料,采用化学还原法制备纳米银(Ag NPs)颗粒,并通过超声波辅助法结合4-羟基香豆素制备4-羟基香豆素-Ag NPs新型复合抑菌材料。利用TEM、XRD、XPS、UV-Vis、FTIR、Zeta电位及理论计算化学等进行表征。以革兰氏阴性菌大肠杆菌(E.coli)、革兰氏阳性菌金黄色葡萄球菌(S. aureus)和耐药菌沙门氏菌(T-Salmonella)为模式菌,研究4-羟基香豆素-Ag的协同抑菌活性及抑菌机制。抑菌性能结果显示:相比Ag NPs,复合材料对E.coli的抑菌效率提高了62.5%,对S. aureus提高了37.5%,对T-Salmonella提高了44.4%。复合材料在浓度为150 μg/mL时,60 min内时对测试菌的抑菌率均可达99.9%。抑菌机制表明:复合材料可显著破坏细菌细胞壁,并进入细菌内部抑制细菌呼吸系统。该材料不仅具备单独无机抗菌剂和有机抗菌剂特有杀菌特性,两者协同且具备更强的抑菌活性,同时还可解决细菌耐药性问题,这可为抗生素的改性和新型抑菌剂的开发提供科学依据。Abstract: The abuse of antibiotics leads to the emergence of a large number of drug-resistant bacteria, which poses a serious threat to health and social economy. Therefore, it is urgent to develop new antibacterial agents to solve the problem of bacterial drug resistance. In this paper, silver trifluoroacetate (CF3COOAg) as raw material, using chemical reduction method to prepare nano-silver (Ag NPs) particles, and ultrasonic assisted coordination with 4-hydroxycoumarin to prepare 4-hydroxycoumarin-Ag NPs new composite antibacterial material. TEM, XRD, XPS, UV-Vis, FTIR, Zeta potential and theoretical calculation were used for characterization. Gram-negative bacteria E.coli, Gram-positive bacteria S. aureus and drug-resistant bacteria T-Salmonella were used as model bacteria to study the synergistic bacteriostatic activity and bacteriostatic mechanism of 4-hydroxycoumarin-Ag. Compared with Ag NPs, the antibacterial efficiency of the composite material for E.coli, S. aureus and T-Salmonella has been improved 62.5%, 37.5% and 44.4% respectively. When the concentration of the composite material was 150 μg/mL, the bacteriostatic rate of the tested bacteria could reach 99.9% within 60 min. The inhibition mechanism showed that the composite material could significantly destroy the bacterial cell wall and enter the bacterial interior to inhibit the bacterial respiratory system. The material not only has the unique bactericidal properties of inorganic antibacterial agent and organic antibacterial agent, but also has stronger antibacterial activity and can solve the problem of bacterial resistance, which can provide scientific basis for the modification of antibiotics and the development of new antibacterial agent.
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Key words:
- synergistic antibacterial /
- Ag NPs /
- 4-hydroxycoumarin /
- antibacterial mechanism /
- coordination
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图 1 纳米银颗粒(Ag NPs)及 4-羟基香豆素-Ag复合材料的TEM图像和粒径分布图((a), (b));(c) Ag NPs和复合材料的XRD图谱;(d) Ag NPs的XPS图谱;(e) Ag NPs、4-羟基香豆素及复合材料的UV-Vis吸收光谱;(f) Ag NPs、4-羟基香豆素及复合材料的FTIR图谱;(g) Ag NPs与4-羟基香豆素结合理论计算分析图
Figure 1. TEM images and particle size distribution map of Ag NPs and 4-hydroxycoumarin-Ag composite ((a), (b)); (c) XRD patterns of Ag NPs and composite materials; (d) XPS spectra of Ag NPs; (e) UV-Vis absorption spectra of Ag NPs, 4-hydroxycoumarin and composite materials; (f) FTIR spectra of Ag NPs, 4-hydroxycoumarin and composite materials; (g) Theoretical calculation and analysis diagram of Ag NPs combined with 4-hydroxycoumarin
图 2 不同材料对E.coli (a)、S.aureus (b)和T-Salmonella (c)的滤纸片扩散照片:浓度为35.5、75、150、300 μg/mL的不同抑菌材料(4-羟基香豆素负载量为5wt%)对E.coli ((a1)~(a4))、S.aureus ((b1)~(b4))及T-Salmonella的抑菌结果((c1)~(c4));E.coli (d)、S.aureus (e)、T-Salmonella (f)的抑菌圈直径随浓度变化曲线
A, B, C and D—Dimethyl sulfoxide, Ag NPs, 4-hydroxycoumarin and composite materials, respectively
Figure 2. Different materials for E.coli (a), S.aureus (b) and T-Salmonella (c) filter paper spread photos: Bacteriostatic results of E.coli of different bacteriostatic materials (The loading of 4-hydroxycoumarin was 5wt%) ((a1)-(a4)), S.aureus ((b1)-(b4) and T-Salmonella ((c1)-(c4)) with concentrations of 35.5, 75, 150 and 300 μg/mL; E.coli (d), S.aureus (e) and T-Salmonella (f) bacteriostatic rings diameter changes with concentration, respectively
图 3 4-羟基香豆素-Ag复合材料对E.coli (a)、S.aureus (b)及T-Salmonella (c)的菌落计数分布图;(d)复合材料的时间-杀菌曲线图;(e) 3种测试菌在不同时间的抑菌率比较
Figure 3. Photo of colony count of 4-hydroxycoumarin-Ag composite for E.coli (a), S.aureus (b) and T-Salmonella (c); (d) Time-germicidal curves of the composite; (e) Comparison of the bacteriostatic rates of the three tested bacteria at different time
图 6 4-羟基香豆素-Ag复合材料对E.coli (a)、S.aureus (b)、T-Salmonella (c)的微量热分析及复合材料毒理性实验结果分析(d)
GDC-0941—Pictilisib; IC50—Half-inhibitory concentration
Figure 6. Microthermal analysis of 4-hydroxycoumarin-Ag composite for E.coli (a), S.aureus (b) and T-Salmonella (c) and analysis of toxicity test results of composites (d)
表 1 溶剂、Ag NPs、4-羟基香豆素、4-羟基香豆素-Ag对E.coli 、S.aureus 和T-Salmonella的抑菌圈尺寸
Table 1. Size of bacteriostasis circles for E.coli, S.aureus and T-Salmonella of solvent, Ag NPs, 4-hydroxycoumarin and 4-hydroxycoumarin-Ag
Bacterial Concentration/(μg·mL−1) Inhibition zone/cm Dimethyl sulfoxide Ag 4-hydroxycoumarin 4-hydroxycoumarin-Ag E. coli 35.5 0.6 0.60 0.60 0.70 75.0 0.6 0.70 0.60 1.00 150.0 0.6 0.90 0.65 1.50 300.0 0.6 1.20 0.70 1.95 S. aureus 35.5 0.6 0.60 0.60 0.65 75.0 0.6 0.65 0.60 0.90 150.0 0.6 0.75 0.65 1.25 300.0 0.6 1.20 0.85 1.62 T-Salmonella 35.5 0.6 0.60 0.60 0.60 75.0 0.6 0.65 0.65 0.80 150.0 0.6 0.70 0.65 1.10 300.0 0.6 0.90 0.70 1.30 -
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