School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daehak-ro 80, Buk-gu, Daegu 702-701, Republic of Korea.
College of Pharmacy, Chosun University, 375 Seosuk-dong, Dong-gu, Gwangju 501-759, Republic of Korea.
Biochem Biophys Res Commun. 2014 Jan 31;444(1):30-5. doi: 10.1016/j.bbrc.2014.01.019. Epub 2014 Jan 14.
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.
钩吻叶苷从忍冬科忍冬属植物获得，其生物活性尚未见报道。本文研究了钩吻叶苷对致病性真菌的抗真菌活性及其作用模式。钩吻叶苷对人红细胞几乎没有溶血作用，但具有抗真菌活性。为了了解其抗真菌机制，进行了膜研究。使用两种染料，3,3'-二丙基噻吖啶碘化物[DiSC3(5)]和碘化丙啶，证实了钩吻叶苷导致膜去极化和通透性增加。此外，通过合成模型膜（包裹钙黄绿素的大单室囊泡[LUVs]）以及观察不同大小的荧光染料（如钙黄绿素、FD4 和 FD10）进入真菌细胞的情况，证明了膜活性机制。膜活性作用是形成孔径为 1.4 至 2.3nm 的孔的作用。最后，三维（3D）流式细胞术分析显示真菌细胞因膜损伤而收缩。总之，本研究表明钩吻叶苷通过破坏膜的机制发挥抗真菌活性。
