Antifungal effect and mode of action of glochidioboside against Candida albicans membranes |
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Authors: | Heejeong Lee Hyemin Choi Hae Ju Ko Eun-Rhan Woo Dong Gun Lee |
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Institution: | 1. School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, Republic of Korea;2. College of Pharmacy, Chosun University, 375 Seosuk-dong, Dong-gu, Gwangju 501-759, Republic of Korea |
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Abstract: | Glochidioboside was obtained from Sambucus williamsii and its biological effect has not been reported. Its antifungal activity against pathogenic fungi and the mode of action involved in its effect were examined. Glochidioboside exerted antifungal effect with almost no hemolytic effect against human erythrocytes. To understand its antifungal mechanisms, membrane studies were done. Using two dyes, 3,3′-dipropylthiacarbocyanine iodide DiSC3(5)] and propidium iodide, membrane depolarization and permeabilization by glochidioboside were confirmed. Furthermore, the membrane-active mechanism was proven by synthesizing a model membrane, calcein-encapsulating large unilamellar vesicles (LUVs), and also by observing the influx of different sized fluorescent dyes, such as calcein, FD4 and FD10, into the fungal cells. The membrane-active action was pore-forming action with radii between 1.4 and 2.3 nm. Finally, three dimensional (3D) flow cytometric analysis showed the shrinkage of the fungal cells from the membrane damage. In conclusion, this study suggests that glochidioboside exerts an antifungal activity through a membrane-disruptive mechanism. |
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Keywords: | MeOH methanol CH2Cl2 dichloromethane EtOAc ethyl acetate n-BuOH n-butanol ATCC American Type Culture Collection KCTC Korean Collection for Type Cultures KRIBB Korea Research Institute of Bioscience and Biotechnology MIC minimum inhibitory concentration CLSI Clinical and Laboratory Standards Institute MTT 3-(4 5-dimethyl-2-thiazolyl)-2 5-diphenyl-2H-tetrazolium bromide PBS phosphate buffered saline DiSC3(5) 3 3&prime -dipropylthiacarbocyanine iodide LUV large unilamellar vesicle PC phosphatidylcholine PE phosphatidylethanolamine PI phosphatidylinositol 3D three dimensional FSC forward scatter SSC side scatter |
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