Mutation Research 723 (2011) 65–71 Contents lists available at ScienceDirect Mutation Research/Genetic Toxicology and Environmental Mutagenesis journal homepage: www.elsevier.com/locate/gentox Community address: www.elsevier.com/locate/mutres Dietary supplementation with whey protein and ginseng extract counteracts oxidative stress and DNA damage in rats fed an aflatoxin-contaminated diet Sekena H. Abdel-Aziem a , Aziza M. Hassan a , Mosaad A. Abdel-Wahhab b,∗ a Cell Biology Department, National Research Center, Dokki, Cairo, Egypt b Food Toxicology & Contaminants Department, National Research Center, Dokki, Cairo, Egypt article info Article history: Received 1 May 2010 Received in revised form 14 March 2011 Accepted 13 April 2011 Available online 21 April 2011 Keywords: Ginseng Whey-protein concentrate Aflatoxin DNA damage Gene expression Oxidative stress Liver Testis abstract Aflatoxins (AF) are among the most potent naturally occurring carcinogens and aflatoxin-B1 (AFB 1 ) is classified as a group-1 carcinogen. Since the ingestion of aflatoxins-contaminated food is associated with several liver diseases, the aim of the present study was to evaluate whether AF-induced damage in rats can be counteracted by feeding with whey-protein concentrates (WPC) and Korean ginseng extract (KGE). Eighty male Sprague-Dawley rats were divided into eight equal groups and treated daily for 30 days as follows: a control group (fed an AF-free diet), a group fed ad libitum an AF-contaminated diet (2.5 mg/kg diet), a group treated orally with WPC (0.5 ml/rat/day), a group treated orally with KGE (20 mg/kg body weight), a group treated orally with WPC + KGE, and three groups that were fed the AF-contaminated diet and were treated orally with WPC, KGE or WPC + KGE, respectively. Throughout the experimental period, animals received WPC or KGE during the consumption of their respective diet. Bone-marrow micronucleus formation, DNA fragmentation, fatty-acid synthesis (FAS) and phospholipid- hydroperoxide-glutathione-peroxidase (PHGPx) mRNA expression, and oxidative stress were assayed in liver and testis. The results indicated that ingestion of aflatoxin resulted in a significant increase in micronucleated normochromatic erythrocytes (Mn-NCE) in bone marrow, DNA fragmentation, FAS mRNA expression and lipid peroxidation in both organs, and a significant decrease in micronucleated poly- chromatic erythrocytes/micronucleated normochromatic erythrocytes (PCE/NCE) ratio in bone marrow, PHGPx gene expression and GSH in liver and testis. Treatments with WPC and/or KGE had a significant effect on Mn-NCE or the PCE/NCE ratio in bone marrow. However, KGE or KGE + WPC increased PHGPx gene expression and GSH in testis accompanied with a significant decrease in lipid peroxidation in liver and testis and FAS-mRNA expression in liver. WPC, KGE or WPC + KGE treatments combined with expo- sure to an AF-contaminated diet restored all the test parameters towards control values, although they did not fully reverse the effects of the aflatoxins. It is suggested that the genotoxicity of aflatoxins can be in part prevented by dietary supplementation with WPC, KGE or their combination. © 2011 Elsevier B.V. All rights reserved. 1. Introduction Aflatoxins (AF) are potent mycotoxins produced by fungi of the Aspergillus group. They have a wide range of biological activities, such as oxidative stress [1,2], acute toxicity, teratogenicity, muta- genicity and carcinogenicity [3–5]. Aflatoxin-B 1 is metabolically bio-transformed in the liver into a highly reactive electrophilic compound that interacts with cellular macromolecules [3]. It has been previously reported that drug-metabolizing enzymes (phase-I and phase-II enzymes) and AFB 1 -adduct formation can be modu- lated by natural constituents of the diet, nutrients and xenobiotics [6]. ∗ Corresponding author. Tel.: +20 2 2283 1943; fax: +20 2 3337 0931. E-mail address: mosaad abdelwahhab@yahoo.com (M.A. Abdel-Wahhab). Panax ginseng C.A. Meyer is an herbal root that has been used for more than 2000 years throughout countries of the Far-East, including China, Japan, and Korea. Its beneficial effects have been analyzed by extensive preclinical and epidemio- logical studies [7,8]. Recently, 20-O-(h-d-glucopyranosyl)-20(S)- protopanaxadiol (IH-901), a novel ginseng saponin metabolite formed from ginsenosides Rb1, Rb2, and Rc was isolated and purified after giving ginseng extract orally to humans and rats [9]. IH-901 has been shown to enhance the efficacy of anticancer drugs in cancer cell lines that were previously resis- tant to such drugs [10]. IH-901 also exhibits anti-genotoxic and anti-clastogenic activity in rats concurrently treated with benzo(a)pyrene [11], and it induces apoptosis [12]. Moreover, several studies indicated that apoptosis is characterized by DNA fragmentation, chromatin condensation, membrane blebbing, cell shrinkage and disassembly into membrane-enclosed vesicles [13]. 1383-5718/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.mrgentox.2011.04.007