4-羟基香豆素-Ag复合材料的协同抑菌性能

梁犇; 吴娟娟; 郑锦丽; 宋凤敏; 徐进; 乔艳明; 郭少波; 史娟

doi:10.13801/j.cnki.fhclxb.20221019.003

4-羟基香豆素-Ag复合材料的协同抑菌性能

doi: 10.13801/j.cnki.fhclxb.20221019.003

梁犇¹,
吴娟娟¹,
郑锦丽¹,
宋凤敏¹,
徐进³,
乔艳明²,
郭少波^1, ,,
史娟^1, ,

1.
陕西理工大学　化学与环境科学学院，汉中 723001
2.
陕西理工大学　生物科学与工程学院，汉中 723001
3.
镇巴县科技进步促进中心，汉中 723001

基金项目: 陕西省科技厅项目(2020 ZDLGY11-02；2021 SF-382；2021 JQ-752)；陕西理工大学秦巴生物资源与生态环境省部共建国家重点实验室项目(SXJ-2106)；陕西省教育厅项目(22 JK0320)；陕西理工大学科研一般项目(SLGKYXM2208)

详细信息

通讯作者:
郭少波，硕士，讲师，研究方向为生物材料　E-mail: 1403699360@qq.com；

史娟，博士，教授，硕士生导师，研究方向为天然产物提取和有机合成　E-mail: 446269824@qq.com

中图分类号: O62；TB333
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出版历程
- 收稿日期: 2022-08-22
- 修回日期: 2022-09-19
- 录用日期: 2022-09-25
- 网络出版日期: 2022-10-20
- 刊出日期: 2023-08-15

Synergistic bacteriostatic properties of 4-hydroxycoumarin-Ag composite

1.
School of Chemistry and Environmental Sciences, Shaanxi University of Technology, Hanzhong 723001, China
2.
School of Biological Sciences and Engineering, Shaanxi University of Technology, Hanzhong 723001, China
3.
Zhenba County Science and Technology Progress Promotion Center, Hanzhong 723001, China

Funds: Project of Science and Technology Department of Shaanxi Province (2020 ZDLGY11-02; 2021 SF-382; 2021 JQ-752); State Key Laboratory Project of Qinba Biological Resources and Ecological Environment, Shaanxi University of Technology (SXJ-2106); Project of Shaanxi Education Department (22 JK0320); General Scientific Research Project of Shaanxi University of Technology (SLGKYXM2208)

摘要

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。
- 协同抑菌 /
- 纳米银 /
- 4-羟基香豆素 /
- 抑菌机制 /
- 配位
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.
- synergistic antibacterial /
- Ag NPs /
- 4-hydroxycoumarin /
- antibacterial mechanism /
- coordination

HTML全文

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

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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

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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

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图 4 4-羟基香豆素-Ag复合材料与E.coli、S.aureus及T-Salmonella作用不同时间的Zeta电位值

Figure 4. Zeta potential diagrams of the 4-hydroxycoumarin-Ag composite reacted with E.coli, S.aureus and T-Salmonella for different time

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图 5 4-羟基香豆素-Ag复合材料对E.coli (b)、S.aureus (d)及T-Salmonella (f)的碘化丙啶(PI)染色照片；((a), (c), (e))对应的纯菌对照效果图

Figure 5. Color propyl iodide (PI) photos of 4-hydroxycoumarin-Ag composite for E.coli (b), S.aureus (d) and T-Salmonella (f); ((a), (c), (e)) Corresponding pure bacteria control renderings

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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)

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图 7 4-羟基香豆素-Ag复合材料抑菌机制

Figure 7. Diagram of antibacterial mechanism of 4-hydroxycoumarin-Ag composite

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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
Bacterial	Concentration/(μg·mL⁻¹)	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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