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Amentoflavone Stimulates Mitochondrial Dysfunction and Induces Apoptotic Cell Death in Candida albicans

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Abstract

Amentoflavone was isolated from an ethyl acetate extract of the whole plant of Selaginella tamariscina. It is a traditional herb for the therapy of chronic trachitis and exhibits some anti-tumor activity. Previously, we confirmed the antifungal effects of amentoflavone. The objective of this study was to investigate the antifungal mechanism(s) of amentoflavone, such as mitochondria-mediated apoptotic cell death. The cells that were treated with amentoflavone exhibited a series of cellular changes that were consistent with apoptosis: externalization of phosphatidylserine, DNA and nuclear fragmentation, accumulation of intracellular reactive oxygen species (ROS) and hydroxyl radicals, and activation of metacaspase. In addition, diagnostic markers of apoptosis, including the reduction of mitochondrial inner-membrane potential and the release of cytochrome c from mitochondria, were observed. These phenomena are important changes in mitochondria-mediated apoptosis. Furthermore, the effect of thiourea as hydroxyl radical scavenger on amentoflavone-induced apoptosis was evaluated. A hydroxyl radical is a more active ROS species. Mitochondrial dysfunction was inhibited, which was indicated by decreased levels of intracellular hydroxyl radicals. Taken together, our results present the first evidence that amentoflavone induces apoptosis in C. albicans cells and is associated with the mitochondrial dysfunction. Besides, amentoflavone-induced hydroxyl radicals may play a significant role in mitochondria-mediated apoptosis.

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Abbreviations

ROS:

Reactive oxygen species

DHR-123:

Dihydrorhodamine

HPF:

2-[6-(4′-hydroxy) phenoxy-3H-xanthen-3-on-9-yl] benzoic acid

DAPI:

4′-6-Diamidino-2-phenylindole

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

DiOC6(3):

3,3′-Dihexyloxacarbocyanine iodide

AMB:

Amphotericin B

PI:

Propidium iodide

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Acknowledgments

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2011-0000915).

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Authors and Affiliations

  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

    In-sok Hwang, Juneyoung Lee & Dong Gun Lee

  2. College of Pharmacy, Chosun University, 375 Seosuk-Dong, Dong-gu, Gwangju, 501-759, Republic of Korea

    Hong-Guang Jin & Eun-Rhan Woo

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  1. In-sok Hwang
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Correspondence to Dong Gun Lee.

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Hwang, Is., Lee, J., Jin, HG. et al. Amentoflavone Stimulates Mitochondrial Dysfunction and Induces Apoptotic Cell Death in Candida albicans . Mycopathologia 173, 207–218 (2012). https://doi.org/10.1007/s11046-011-9503-x

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