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Antimicrobial Agents and Chemotherapy, October 2002, p. 3113-3117, Vol. 46, No. 10
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.10.3113-3117.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Endogenous Reactive Oxygen Species Is an Important Mediator of Miconazole Antifungal Effect

Daisuke Kobayashi,1,2 Kei Kondo,2 Nobuyuki Uehara,2 Seiko Otokozawa,2 Naoki Tsuji,1,2 Atsuhito Yagihashi,1,2 and Naoki Watanabe1,2*

Department of Clinical Laboratory Medicine,1 Division of Laboratory Diagnosis, Sapporo Medical University School of Medicine, Sapporo, Japan2

Received 25 March 2002/ Returned for modification 7 May 2002/ Accepted 26 June 2002

We investigated the significance of endogenous reactive oxygen species (ROS) produced by fungi treated with miconazole. ROS production in Candida albicans was measured by a real-time fluorogenic assay. The level of ROS production was increased by miconazole at the MIC (0.125 µg/ml) and was enhanced further in a dose-dependent manner, with a fourfold increase detected when miconazole was used at 12.5 µg/ml. This increase in the level of ROS production was completely inhibited by pyrrolidinedithiocarbamate (PDTC), an antioxidant, at 10 µM. In a colony formation assay, the decrease in cell viability associated with miconazole treatment was significantly prevented by addition of PDTC. Moreover, the level of ROS production by 10 clinical isolates of Candida species was inversely correlated with the miconazole MIC (r = -0.8818; P < 0.01). These results indicate that ROS production is important to the antifungal activity of miconazole.

* Corresponding author. Mailing address: Department of Clinical Laboratory Medicine, Sapporo Medical University School of Medicine, South-1, West-16, Chuo-ku, Sapporo 060-0061, Japan. Phone: 81-11-611-2111. Fax: 81-11-622-7502. E-mail: watanabn{at}sapmed.ac.jp.

Antimicrobial Agents and Chemotherapy, October 2002, p. 3113-3117, Vol. 46, No. 10
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.10.3113-3117.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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