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基于FT-IR研究环丙沙星和白花丹素对金黄色葡萄球菌的抗菌作用
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Abstract:
目的：本研究旨在探讨环丙沙星(CIP)与白花丹素(PLU)对不同生长阶段的金黄色葡萄球菌(SA)的动态杀菌过程及其细胞结构改变。方法：利用剂量–反应曲线和时间杀伤曲线评估两种药物在指数期和稳定期的杀菌动力学；采用傅里叶变换红外光谱(FT-IR)监测细菌在药物作用后细菌细胞膜、蛋白质、核酸等结构及胞外多糖(EPS)的变化；利用主成分分析(PCA)和支持向量机(SVM)模型对光谱数据进行特征提取和分类验证；采用透射电子显微镜(TEM)技术观察药物处理前后SA的超微结构变化。结果：CIP对指数期SA杀伤迅速，而PLU在稳定期表现出更强的抑制作用，两药联用在不同生长阶段均表现出相加或协同效应。光谱分析证实，CIP主要破坏细胞膜完整性并在作用后期损伤核酸结构；PLU虽对膜破坏有限，但显著改变蛋白质空间构象，单独或与CIP联合显著减少了EPS分泌。PCA显示各处理组间差异显著，SVM模型分类准确率最高达100%。TEM显示CIP和PLU均破坏了SA的细胞膜。结论：本研究展示了CIP与PLU在金黄色葡萄球菌上的多靶点联合作用(膜、蛋白、核酸及EPS等)，为应对耐药性贡献了潜在策略，为时序性联合用药提供了依据，并展示了FT-IR在监测抗菌动态过程中的潜在应用价值。
Objective: This study aims to investigate the dynamic bactericidal process and associated changes in cell structure of Staphylococcus aureus (SA) at different growth stages following treatment with ciprofloxacin (CIP) and plumbagin (PLU). Methods: Dose-response curves and time-kill curves were utilised to evaluate the bactericidal kinetics of the two drugs during the log and stationary phases; Fourier transform infrared spectroscopy (FT-IR) was employed to monitor changes in bacterial cell membranes, proteins, nucleic acids and extracellular polysaccharides (EPS) following drug exposure; principal component analysis (PCA) and support vector machine (SVM) models were used for feature extraction and classification validation of the spectral data; The ultrastructural changes in SA before and after drug treatment were observed using transmission electron microscopy (TEM). Results: CIP rapidly killed SA during the log phase, whilst PLU exhibited stronger inhibitory effects during the stationary phase; the combination of the two drugs demonstrated additive or synergistic effects across different growth stages. Spectral analysis confirmed that CIP primarily disrupted cell membrane integrity and damaged nucleic acid structures in the later stages of action; although PLU caused limited membrane disruption, it significantly altered the spatial conformation of proteins and, either alone or in combination with CIP, significantly reduced EPS secretion. PCA analysis revealed significant differences between treatment groups, with the SVM model achieving a classification accuracy of up to 100%. Conclusion: This study demonstrates the multi-targeted synergistic effects of CIP and PLU against Staphylococcus aureus (acting on membranes, proteins, nucleic acids, and EPS, among others), offering a potential strategy for combating antimicrobial resistance, providing a basis for sequential combination therapy, and highlighting
| [1] | Cacace, E., Kim, V., Varik, V., Knopp, M., Tietgen, M., Brauer-Nikonow, A., et al. (2023) Systematic Analysis of Drug Combinations against Gram-Positive Bacteria. Nature Microbiology, 8, 2196-2212. https://doi.org/10.1038/s41564-023-01486-9 |
| [2] | Liu, X., Xu, W., Feng, J., Wang, Y., Li, K., Chen, Y., et al. (2025) Adoptive Cell Transfer of Piezo-Activated Macrophage Rescues Immunosuppressed Rodents from Life-Threating Bacterial Infections. Nature Communications, 16, Article No. 1363. https://doi.org/10.1038/s41467-025-56460-2 |
| [3] | Mlynarczyk-Bonikowska, B. and Rudnicka, L. (2025) The Pathogenicity Mechanisms of Staphylococcus aureus. International Journal of Molecular Sciences, 26, Article No. 11803. https://doi.org/10.3390/ijms262411803 |
| [4] | Kaul, G., Akhir, A., Shukla, M., Shafi, H., Akunuri, R., Pawar, G., et al. (2023) Oxiconazole Potentiates Gentamicin against Gentamicin-Resistant Staphylococcus aureus in Vitro and in Vivo. Microbiology Spectrum, 11, Article ID: 05031-22. https://doi.org/10.1128/spectrum.05031-22 |
| [5] | AL-Harbi, A.I., AL-Mohammadi, N.A., AL-Hubayshi, N.E. and AL-Mehayawi, S.S. (2024) Prevention and Control towards Hospital-Acquired Infections, Mixed-Methods Systemic Review. Global Journal of Health Science, 16, Article No. 48. https://doi.org/10.5539/gjhs.v16n6p48 |
| [6] | Silva-Santana, G. (2025) Staphylococcus aureus: Dynamics of Pathogenicity and Antimicrobial-Resistance in Hospital and Community Environments—Comprehensive Overview. Research in Microbiology, 176, Article ID: 104267. https://doi.org/10.1016/j.resmic.2025.104267 |
| [7] | Jia, Y. and Zhao, L. (2021) The Antibacterial Activity of Fluoroquinolone Derivatives: An Update (2018-2021). European Journal of Medicinal Chemistry, 224, Article ID: 113741. https://doi.org/10.1016/j.ejmech.2021.113741 |
| [8] | Saeful, A., Fuza Khoirun, N., Yosi, S., et al. (2025) Kajian Kimia Medisinal Ciprofloxacin: Mekanisme Kerja, Antibakteri, dan Pola Resistensi Bakteri. Jurnal Ilmiah Kedokteran dan Kesehatan, 4, 121-131. https://doi.org/10.55606/klinik.v4i2.3923 |
| [9] | Yu, X.H., Hao, Z.H., Liu, P.L., Liu, M.M., Zhao, L.L. and Zhao, X. (2022) Increased Expression of Efflux Pump norA Drives the Rapid Evolutionary Trajectory from Tolerance to Resistance against Ciprofloxacin in Staphylococcus aureus. Antimicrobial Agents and Chemotherapy, 66, Article ID: 00594-22. https://doi.org/10.1128/aac.00594-22 |
| [10] | Hwang, J., Barman, S., Gao, R., Yang, X., O’Malley, A., Nagarkatti, P., et al. (2023) Membrane‐Active Metallopolymers: Repurposing and Rehabilitating Antibiotics to Gram‐Negative Superbugs. Advanced Healthcare Materials, 12, Article ID: 2301764. https://doi.org/10.1002/adhm.202301764 |
| [11] | Si, Z., Pethe, K. and Chan-Park, M.B. (2023) Chemical Basis of Combination Therapy to Combat Antibiotic Resistance. JACS Au, 3, 276-292. https://doi.org/10.1021/jacsau.2c00532 |
| [12] | Wang, N., Luo, J., Deng, F., Huang, Y. and Zhou, H. (2022) Antibiotic Combination Therapy: A Strategy to Overcome Bacterial Resistance to Aminoglycoside Antibiotics. Frontiers in Pharmacology, 13, Article ID: 839808. https://doi.org/10.3389/fphar.2022.839808 |
| [13] | Zouine, N., Ghachtouli, N.E., Abed, S.E. and Koraichi, S.I. (2024) A Comprehensive Review on Medicinal Plant Extracts as Antibacterial Agents: Factors, Mechanism Insights and Future Prospects. Scientific African, 26, e02395. https://doi.org/10.1016/j.sciaf.2024.e02395 |
| [14] | Guo, L., Li, Y., Feng, J., Li, Y., Liao, Y., Zeng, Q., et al. (2025) Antibacterial Activity and Potential Mechanisms of Plumbagin against Escherichia Coli and Its Application in Milk. Current Research in Food Science, 10, Article ID: 101083. https://doi.org/10.1016/j.crfs.2025.101083 |
| [15] | Janeczko, M., Demchuk, O.M., Strzelecka, D., Kubiński, K. and Masłyk, M. (2016) New Family of Antimicrobial Agents Derived from 1,4-Naphthoquinone. European Journal of Medicinal Chemistry, 124, 1019-1025. https://doi.org/10.1016/j.ejmech.2016.10.034 |
| [16] | Bie, S., Mo, Q., Shi, C., Yuan, H., Li, C., Wu, T., et al. (2024) Interactions of Plumbagin with Five Common Antibiotics against Staphylococcus aureus in Vitro. PLOS ONE, 19, e0297493. https://doi.org/10.1371/journal.pone.0297493 |
| [17] | Periasamy, H., Iswarya, S., Pavithra, N., Senthilnathan, S. and Gnanamani, A. (2019) In Vitro Antibacterial Activity of Plumbagin Isolated from Plumbago zeylanica L. against Methicillin‐Resistant Staphylococcus aureus. Letters in Applied Microbiology, 69, 41-49. https://doi.org/10.1111/lam.13160 |
| [18] | Li, H., Wang, S., Zeng, Q., Chen, C., Lv, X., Ma, M., et al. (2022) Serum Raman Spectroscopy Combined with Multiple Classification Models for Rapid Diagnosis of Breast Cancer. Photodiagnosis and Photodynamic Therapy, 40, Article ID: 103115. https://doi.org/10.1016/j.pdpdt.2022.103115 |
| [19] | Cai, Q., Yuan, R., He, J., Li, M. and Guo, Y. (2021) Predicting HIV Drug Resistance Using Weighted Machine Learning Method at Target Protein Sequence-Level. Molecular Diversity, 25, 1541-1551. https://doi.org/10.1007/s11030-021-10262-y |
| [20] | Wu, C., Huang, H., Huang, S., Chen, I., Liao, S., Chen, C., et al. (2021) Resting-State EEG Signal for Major Depressive Disorder Detection: A Systematic Validation on a Large and Diverse Dataset. Biosensors, 11, Article No. 499. https://doi.org/10.3390/bios11120499 |
| [21] | Bie, S., Yuan, H., Shi, C., Li, C., Lu, M., Yao, Z., et al. (2025) Antibiofilm Activity of Plumbagin against Staphylococcus aureus. Scientific Reports, 15, Article No. 7948. https://doi.org/10.1038/s41598-025-92435-5 |
| [22] | Vásquez, A., Echeverri-Gaviria, S. and Manrique-Moreno, M. (2025) Modulation of Antimicrobial Peptide-Membrane Interactions by Lysyl-Phosphatidylglycerol in Staphylococcus aureus: An FTIR Spectroscopy Study. Scientia Pharmaceutica, 93, Article No. 49. https://doi.org/10.3390/scipharm93040049 |
| [23] | Kamaruzzaman, A.N.A., Tengku Zainal Mulok, T.E., Mohamad Nor, N.H. and Raja Yahya, M.F.Z. (2022) FTIR Spectral Changes in Candida Albicans Biofilm Following Exposure to Antifungals. Malaysian Applied Biology, 51, 57-66. https://doi.org/10.55230/mabjournal.v51i4.11 |
| [24] | Suleiman, M., Abu-Aqil, G., Sharaha, U., Riesenberg, K., Lapidot, I., Salman, A., et al. (2022) Infra-Red Spectroscopy Combined with Machine Learning Algorithms Enables Early Determination of Pseudomonas aeruginosa’s Susceptibility to Antibiotics. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 274, Article ID: 121080. https://doi.org/10.1016/j.saa.2022.121080 |
| [25] | Li, M., Jian, Q., Ye, X., Jing, M., Wu, J., Wu, Z., et al. (2025) Mechanisms of Mepa Overexpression and Membrane Potential Reduction Leading to Ciprofloxacin Heteroresistance in a Staphylococcus aureus Isolate. International Journal of Molecular Sciences, 26, Article No. 2372. https://doi.org/10.3390/ijms26052372 |
| [26] | Shariati, A., Arshadi, M., Khosrojerdi, M.A., Abedinzadeh, M., Ganjalishahi, M., Maleki, A., et al. (2022) The Resistance Mechanisms of Bacteria against Ciprofloxacin and New Approaches for Enhancing the Efficacy of This Antibiotic. Frontiers in Public Health, 10, Article ID: 1025633. https://doi.org/10.3389/fpubh.2022.1025633 |
| [27] | Basavegowda, N. and Baek, K. (2022) Combination Strategies of Different Antimicrobials: An Efficient and Alternative Tool for Pathogen Inactivation. Biomedicines, 10, Article No. 2219. https://doi.org/10.3390/biomedicines10092219 |
| [28] | Manso, T., Lores, M. and de Miguel, T. (2021) Antimicrobial Activity of Polyphenols and Natural Polyphenolic Extracts on Clinical Isolates. Antibiotics, 11, Article No. 46. https://doi.org/10.3390/antibiotics11010046 |
| [29] | Petrocelli, G., Marrazzo, P., Bonsi, L., Facchin, F., Alviano, F. and Canaider, S. (2023) Plumbagin, a Natural Compound with Several Biological Effects and Anti-Inflammatory Properties. Life, 13, Article No. 1303. https://doi.org/10.3390/life13061303 |
| [30] | Huang, A., Su, L., He, W., Zhang, F., Wei, C. and Wang, Y. (2022) Natural Component Plumbagin as a Potential Antibacterial Agent against Streptococcus agalactiae Infection. Journal of Fish Diseases, 45, 815-823. https://doi.org/10.1111/jfd.13606 |
| [31] | Wang, Y., Kong, J., Zhang, X., Liu, Y., Huang, Z., Yuan, L., et al. (2022) Plumbagin Resurrect Colistin Susceptible against Colistin-Resistant Pseudomonas aeruginosa in Vitro and in Vivo. Frontiers in Microbiology, 13, Article ID: 1020652. https://doi.org/10.3389/fmicb.2022.1020652 |
| [32] | Yuan, H., Lu, M., Shi, C., Li, C., Yao, Z., Shang, H., et al. (2025) Plumbagin Disrupts the Mature Biofilm of Staphylococcus aureus. Biofouling, 41, 1132-1144. https://doi.org/10.1080/08927014.2025.2589802 |
