Skip Navigation



Journal of Biochemistry Advance Access published online on September 7, 2006
Journal of Biochemistry, doi:10.1093/jb/mvj187
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
140/4/483    most recent
mvj187v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Andoh, Y.
Right arrow Articles by Taketani, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Andoh, Y.
Right arrow Articles by Taketani, S.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2006 The Japanese Biochemical Society.
Received July 30, 2006
Accepted August 8, 2006

Regular Paper

The Antioxidant Role of a Reagent, 2',7'-Dichlorodihydrofluorescin Diacetate, Detecting Reactive-Oxygen Species and Blocking the Induction of Heme Oxygenase-1 and Preventing Cytotoxicity

Yoshihiro Andoh 1, Atsushi Mizutani 1, Tomoko Ohashi 1, Shosuke Kojo 2, Tetsuro Ishii 3, Yasushi Adachi 4, Susumu Ikehara 4, and Shigeru Taketani 5 *

1 Department of Biotechnology, Kyoto Institute of Technology, Kyoto 606-8585, Japan
2 Department of Food Science, Nara Women's University, Nara 622, Japan; Department of Nutrition, Nara Women's University, Nara 622, Japan
3 Graduate School of Comprehensive Human Sciences, Tsukuba University, Tukuba 305-8575, Japan
4 The First Department of Pathology, Kansai Medical University, Moriguchi, Osaka 570-8506, Japan
5 Department of Biotechnology, Kyoto Institute of Technology, Kyoto 606-8585, Japan; Insect Biomedical Center, Kyoto Institute of Technology, Kyoto 606-8585, Japan

* To whom correspondence should be addressed.
Shigeru Taketani, E-mail: taketani{at}kit.ac.jp


  Abstract

Heme oxygenase-1 (HO-1) degrades heme into biliverdin, iron and CO. The enzyme participates in adaptive and protective responses to oxidative stress and various inflammatory stimuli, and is induced in response to reactive oxygen species (ROS). 2',7'-Dichlorodihydrofluorescin diacetate (DCFH-DA) is a common reagent used to detect ROS by the oxidation of 2',7'-dichlorodihydrofluorescin (DCFH) to fluorescent dichlorodihydrofluorescein. We previously found that rapid oxidation of DCFH occurred with heme-compounds as well as ROS (Ohashi, T., et al. (2002) FEBS Lett. 511, 21-27), and then examined the effect of DCFH-DA on the induction of HO-1 expression by arsenite, cadmium and hemin, which induce oxidative stress and cytotoxicity. We found suppression of the arsenite-, cadmium- and hemin-dependent induction of HO-1 with DCFH-DA. The suppression occurred at the transcriptional level since the promoter activity of the Maf-recognition site of the HO-1 gene decreased with the DCFH-DA treatment. DCFH abolished the phosphorylation of extracellular signal-regulated kinase, the nuclear translocation of a transcriptional activator Nrf2, and cell death. An antioxidant, N-acetylcysteine (NAC), also suppressed the induction by arsenite and cadmium, but not that by hemin, indicating that DCFH blocked a different site in the stress signal pathway from NAC. Considering that the oxidation of DCFH diminishes ROS generated by various stressors, our findings provide a potential strategy for protection of cells from toxic insults using DCFH-like molecules.

Keywords: heme oxygenase-1; ROS; 2',7'-dichlorodihydrofluorescin; Nrf2; cytotoxicity.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.