Paeonia lactiflora Inhibits Cell Wall Synthesis and Triggers Membrane Depolarization in Candida albicans
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Fungal cell walls and cell membranes are the main targets of antifungals. In this study, we report on the antifungal activity of an ethanol extract from Paeonia lactiflora against Candida albicans, showing that the antifungal activity is associated with the synergistic actions of preventing cell wall synthesis, enabling membrane depolarization, and compromising permeability. First, it was shown that the ethanol extract from P. lactiflora was involved in damaging the integrity of cell walls in C. albicans. In isotonic media, cell bursts of C. albicans by the P. lactiflora ethanol extract could be restored, and the minimum inhibitory concentration (MIC) of the P. lactiflora ethanol extract against C. albicans cells increased 4-fold. In addition, synthesis of (1,3)-β-D-glucan polymer was inhibited by 87% and 83% following treatment of C. albicans microsomes with the P. lactiflora ethanol extract at their 1× MIC and 2× MIC, respectively. Second, the ethanol extract from P. lactiflora influenced the function of C. albicans cell membranes. C. albicans cells treated with the P. lactiflora ethanol extract formed red aggregates by staining with a membrane-impermeable dye, propidium iodide. Membrane depolarization manifested as increased fluorescence intensity by staining P. lactiflora-treated C. albicans cells with a membrane-potential marker, DiBAC4(3) ((bis-1,3-dibutylbarbituric acid) trimethine oxonol). Membrane permeability was assessed by crystal violet assay, and C. albicans cells treated with the P. lactiflora ethanol extract exhibited significant uptake of crystal violet in a concentration-dependent manner. The findings suggest that P. lactiflora ethanol extract is a viable and effective candidate for the development of new antifungal agents to treat Candida-associated diseases.
Abstract
Fungal cell walls and cell membranes are the main targets of antifungals. In this study, we report on the antifungal activity of an ethanol extract from Paeonia lactiflora against Candida albicans, showing that the antifungal activity is associated with the synergistic actions of preventing cell wall synthesis, enabling membrane depolarization, and compromising permeability. First, it was shown that the ethanol extract from P. lactiflora was involved in damaging the integrity of cell walls in C. albicans. In isotonic media, cell bursts of C. albicans by the P. lactiflora ethanol extract could be restored, and the minimum inhibitory concentration (MIC) of the P. lactiflora ethanol extract against C. albicans cells increased 4-fold. In addition, synthesis of (1,3)-β-D-glucan polymer was inhibited by 87% and 83% following treatment of C. albicans microsomes with the P. lactiflora ethanol extract at their 1× MIC and 2× MIC, respectively. Second, the ethanol extract from P. lactiflora influenced the function of C. albicans cell membranes. C. albicans cells treated with the P. lactiflora ethanol extract formed red aggregates by staining with a membrane-impermeable dye, propidium iodide. Membrane depolarization manifested as increased fluorescence intensity by staining P. lactiflora-treated C. albicans cells with a membrane-potential marker, DiBAC4(3) ((bis-1,3-dibutylbarbituric acid) trimethine oxonol). Membrane permeability was assessed by crystal violet assay, and C. albicans cells treated with the P. lactiflora ethanol extract exhibited significant uptake of crystal violet in a concentration-dependent manner. The findings suggest that P. lactiflora ethanol extract is a viable and effective candidate for the development of new antifungal agents to treat Candida-associated diseases.
참고문헌 (33)
- Pfaller MA, "Invasive fungal pathogens: current epidemiological trends", Clin. Infect. Dis, 43, pp.S3~S14 (2006)
- Eliopoulos GM, "Antifungal resistance in pathogenic fungi", Clin. Infect. Dis, 35, pp.1073~1080 (2002)
- Jenkinson HF, "Polymicrobial Diseases" (2002), ASM Press
- Lortholary O, "Antifungal prophylaxis during neutropenia and immunodeficiency", Clin. Microbiol. Rev, 10, pp.477~504 (1997)
- Vandeputte P, "Antifungal resistance and new strategies to control fungal infections", Int. J. Microbiol, 2012, pp.1~26 (2011)
- White TC, "Clinical, cellular, and molecular factors that contribute to antifungal drug resistance", Clin. Microbiol. Rev, 11, pp.382~402 (1998)
- Gray KC, "Amphotericin primarily kills yeast by simply binding ergosterol", Proc. Natl. Acad. Sci. USA, 109, pp.2234~2239 (2012)
- Carrillo-Muñoz AJ, "Antifungal agents: mode of action in yeast cells", Rev. Esp. Quimioter, 19, pp.130~139 (2006)
- Kurtz M, "Lipopeptide inhibitors of fungal glucan synthase", J. Med. Vet. Mycol, 35, pp.79~86 (1997)
- Vermes A, "Flucytosine: a review of its pharmacology, clinical indications, pharmacokinetics, toxicity and drug interactions", J. Antimicrob. Chemother, 46, pp.171~179 (2000)
- Carson CF, "Mechanism of action of Melaleuca alternifolia (tea tree) oil on Staphylococcus aureus determined by time-kill, lysis, leakage, and salt tolerance assays and electron microscopy", Antimicrob. Agents Chemother, 46, pp.1914~1920 (2002)
- Shapiro RS, "Regulatory circuitry governing fungal development, drug resistance, and disease", Microbiol. Mol. Biol. Rev, 75, pp.213~267 (2011)
- Cowan MM., "Plant products as antimicrobial agents", Clin. Microbiol. Rev, 12, pp.564~582 (1999)
- Kim CM, "The Encyclopedia of Oriental Herbal Medicine" (1998), Jeongdam Press
- Zhang W, "Mechanisms involved in the therapeutic effects of Paeonia lactiflora Pallas in rheumatoid arthritis", Int. Immunopharmacol, 14, pp.27~31 (2012)
- Clinical and Laboratory Standards Institute, "M27-A3. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts: Approved Standard", Clinical and Laboratory Standards Institute, ~ (2008)
- Liu M, "Colorimetric broth microdilution method for the antifungal screening of plant extracts against yeast", Methods, 42, pp.325~329 (2007)
- Frost DJ, "A wholecell Candida albicans assay for the detection of inhibitors towards fungal cell wall synthesis and assembly", J. Antibiot, 48, pp.306~310 (1995)
- Shedletzky E, "A microtiter-based fluorescence assay for (1,3)-β-glucan synthases", Anal. Biochem, 249, pp.88~93 (1997)
- HEUNG SHICK LEE, "Antifungal Activity of Salvia miltiorrhiza Against Candida albicans Is Associated with the Alteration of Membrane Permeability and (1,3)-β-D-Glucan Synthase Activity", Journal of Microbiology and Biotechnology, 26(3), pp.610~617 (2016), 한국미생물·생명공학회[DOI]
- Vaara M, "Outer membrane permeability barrier disruption by polymixin in polymixin-susceptible and resistant Salmonella typhimurium", Antimicrob. Agents Chemother, 19, pp.578~583 (1981)
- 이흥식, "막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성", 미생물학회지, 50(4), pp.360~367 (2014), 한국미생물학회[DOI]
- Halder S, "Alteration of zeta potential and membrane permeability in bacteria: a study with cationic agents", SpringerPlus, 4, pp.672~ (2015)
- 최인호, "Candida albicans에 대한 桂枝, 白頭翁, 白鮮皮, 白芍藥, 檳？, 茵蔯의 항진균효과", 동의생리병리학회지, 19(3), pp.690~695 (2005), 한의병리학회
- Onishi J, "Discovery of novel antifungal (1,3)-β-D-glucan synthase inhibitors", Antimicrob. Agents Chemother, 44, pp.368~377 (2000)
- Chandra J, "Modification of surface properties of biomaterials influences the ability of Candida albicans to form biofilms", Appl. Environ. Microbiol, 71, pp.8795~8801 (2005)
- Tuomanen E, "The rate of killing of Escherichia coli by β-lactam antibiotics is strictly proportional to the rate of bacterial growth", J. Gen. Microbiol, 132, pp.1297~1304 (1986)
- Ehara M, "Uptake of neutral red by the vacuoles of a green alga, Micrasterias pinnatifida", Plant Cell Physiol, 37, pp.734~741 (1996)
- Viarengo A, "The use of biomarkers in biomonitoring: a 2-tier approach assessing the level of pollutant-induced stress syndrome in sentinel organisms", Comp. Biochem. Physiol. C, 146, pp.281~300 (2007)
- Li SC, "The yeast lysosome-like vacuole:endpoint and crossroads", Biochim. Biophys. Acta, 1793, pp.650~663 (2009)
- Wilson HA, "Lymphocyte membrane potential and Ca2+ sensitivity potassium channels described by oxonol dye fluorescence measurements", J. Cell. Physiol, 125, pp.72~81 (1985)
- Volkov V, "Quantitative description of ion transportvia plasma membrane of yeast and small cells", Front. Plant Sci, 6, pp.1~22 (2015)
- Devi KP, "Eugenol (an essential oil of clove) acts as an antibacterial agent against Salmonella typhi by disrupting the cellular membrane", J. Ethnopharmacol, 130, pp.107~115 (2010)