Auraptene from Ferula szowitsiana Protects Human Peripheral Lymphocytes Against Oxidative Stress Fatemeh Soltani, 1,2 Fatemeh Mosaffa, 1,2 Mehrdad Iranshahi, 1,2 * Gholamreza Karimi, 3,4 Mohammad Malekaneh, 5 Fatemeh Haghighi 6 and Javad Behravan 1,2 1 Biotechnology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran 2 Department of Pharmacognosy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran 3 Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran 4 Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran 5 Department of Clinical Biochemistry, Birjand University of Medical Sciences, Birjand, Iran 6 Department of Pathology, Birjand University of Medical Sciences, Birjand, Iran The antigenotoxicity effects of auraptene on DNA damage in human peripheral lymphocytes were studied using alkaline single cell gel electrophoresis. Auraptene at concentrations of 5, 10, 25, 50, 100, 200 and 400 mM was tested under simultaneous treatment with 25 mM H 2 O 2 . The data are expressed as % tail DNA and com- pared with ascorbic acid at concentrations of 25, 50, 100, 200 and 400 mM. Auraptene significantly reduced the genotoxicity of H 2 O 2 at concentrations higher than 25 mM (p < 0.001). Interestingly, the antigenotoxicity activity of auraptene was higher than ascorbic acid (p < 0.01), however, at some concentrations (25, 50 and 200 mM) there was no significant difference between auraptene and ascorbic acid (p > 0.05). It seems that the significant antigenotoxicity effects of auraptene may be due to the prenyl moiety and also the suppression of superoxide anion (O 2 - ) generation. This study suggests that the antigenotoxic property of auraptene is of great pharma- cological importance and might be beneficial for cancer prevention. Copyright ” 2009 John Wiley & Sons, Ltd. Keywords: antigenotoxicity; comet assay; auraptene. INTRODUCTION Oxidative stress is known to play an important role in the etiology of several human diseases such as cancer, atherosclerosis, arthritis and aging (Bonomini et al., 2008; Ishii, 2007; Laviano et al., 2007; Seven et al., 2008). Oxidative stress can occur through the overproduction of reactive oxygen species (ROS). Reactive oxygen species are formed during normal cell aerobic respira- tion (Barzilai and Yamamoto, 2004). They may not only have an impact on DNA damage but also influence DNA messengers to modulate DNA replication and the cell cycle (Hu et al., 1995). Thus, to eliminate oxidative stress such as reactive oxygen species in the body, many researchers have been attempting to develop antioxi- dant agents (Baratta and Ruberto, 2000; Kogure et al., 2004; Torres et al., 2006; Yen et al., 2002). There is some evidence that some plant-derived chemicals have pro- tective effects towards DNA damage due to oxidative stresses (Glei and Pool-Zobel, 2006; Plazar et al., 2008). Coumarins are a large class of natural derivatives mainly found in the families Umbelliferae and Rutaceae (Curini et al., 2006). Auraptene, 7-geranloxy- coumarin (Fig. 1), is synthesized by various plant species such as Ferula and Citrus (Iranshahi et al., 2007; Ju-Ichi, 2005). It is endowed with interesting medicinal proper- ties including antileishmanial, antihelicobacter and anti- oxidant activities (Iranshahi et al., 2007; Murakami and Ohigashi, 2006; Takeda et al., 2007). Auraptene has been reported to be an effective inhibitor of chemical carcinogenesis in some rodent models (Kohno et al., 2006). In addition to an array of biological effects, a significant cancer chemoprevention activity of aurap- tene was reported (Iranshahi et al., 2008; Ju-Ichi, 2005). Also, it can suppress superoxide anion (O 2 - ) generation from inflammatory leucocytes in in vitro experiments (Murakami et al., 2000). Despite the interesting biologi- cal activities of auraptene, there is no published report about the antigenotoxicity effect of this compound. Recently, the antigenotoxicity effect of umbellip- renin, 7-farnesyloxycoumarin (Fig. 1) which has a struc- ture close to auraptene, was reported (Soltani et al., 2008). In view of the fact that the only difference between umbelliprenin and auraptene structures is the higher length of the prenyl chain in umbelliprenin and due to the lack of information about its antigenotoxic- ity, this paper was addressed to evaluate the antigeno- toxicity effects of auraptene against DNA damage induced by hydrogen peroxide in human lymphocytes. The study used the most common technique, the comet assay (Rojas et al., 1999) which is a rapid, simple, sensi- tive and reliable biochemical method for evaluating DNA damage in cells or in tissues. Furthermore, the antigenotoxicity effect of auraptene was compared with those reported for umbelliprenin. Received 27 January 2009 Revised 11 March 2009 Copyright © 2009 John Wiley & Sons, Ltd. Accepted 24 March 2009 PHYTOTHERAPY RESEARCH Phytother. Res. 24: 85–89 (2010) Published online 14 May 2009 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ptr.2874 * Correspondence to: Dr Mehrdad Iranshahi, Department of Pharmacog- nosy, Biotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences (MUMS), P.O. Box: 91775-1365, Mashhad, Iran. E-mail: Iranshahim@mums.ac.ir