Aziza M. Hassan

Dietary supplementation with whey protein and ginseng extract counteracts oxidative stress and DNA damage in rats fed an aflatoxin-contaminated diet.

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 (2.5mg/kg diet), a group treated orally with WPC (0.5ml/rat/day), a group treated orally with KGE (20mg/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 polychromatic 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 exposure 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.

Quercetin inhibits the cytotoxicity and oxidative stress in liver of rats fed aflatoxin-contaminated diet

This study was conducted to evaluate the protective role of quercetin (Q) against the cytotoxicity, DNA damage and oxidative stress in rats fed aflatoxin (AFs)-contaminated diet. Female Sprague–Dawley rats were divided into six groups and treated for 21 days as follows: the control group; the group fed AFs-contaminated diet (1.4 mg/kg diet); the groups treated orally with Q at low or high dose (50 and 100 mg/kg b.w.) and the groups AFs-contaminated diet plus low or high dose of Q. At the end of experiment, blood and liver samples were collected for biochemical, histological, histochemical and genetic analyses. The results indicated that animal fed AFs-contaminated diet showed significant increase in serum biochemical parameters, oxidative stress markers and DNA fragmentation accompanied with significant decrease in total proteins, GPX, SOD, DNA and RNA content and fatty acid synthase (Fas) and TNFα gene expression in the liver tissue. Q at the two tested doses succeeded to normalize the biochemical parameters, improved the content of nucleic acids in hepatic tissues, the gene expression, the histopathological and histochemical picture of the liver. It could be concluded that Q has a potential antioxidant activity, a protective action and regulated the alteration of genes expression induced by AFs.

Aquilegia vulgaris L. extract counteracts oxidative stress and cytotoxicity of fumonisin in rats

Exposure to fumonisins (FB) is known to have toxic and carcinogenic effects in different animal species, and to express toxicity in cells via the induction of oxidative stress. The aim of the current study was to evaluate the protective effects of the ethanol extract of Aquilegia vulgaris L. against the oxidative stress and the genotoxicity using micronucleus assay and random amplified polymorphism DNA (RAPD-PCR) in FB-treated rats. Sixty mature female Sprague-Dawley were divided into six treatment groups and treated for 4 weeks as follow: the control group, the group fed FB-contaminated diet (200 mg/kg diet), the groups treated orally with the extract (5 and 10 mg/kg bw) and the groups fed FB-contaminated diet and treated with the extract at the two doses. The results showed that treatment with FB alone disturbed lipid profile in serum, increases Sa/So ratio, induces micronucleated polychromatic erythrocytes (Mn-PCEs) in bone marrow, increases DNA and RNA in liver accompanied with significant changes in histological picture The extract alone at the two tested doses did not induce any significant changes in the biochemical or histological picture. The combined treatment showed significant improvements in all biochemical, cytogenetic parameters tested and histological pictures in the liver tissues. Moreover, this improvement was more pronounced in the group received the high dose of the extract. It could be concluded that the ethanol extract of A. vulgaris induced its protective effect via the increase in the antioxidant capacity, inhibition of lipid peroxidation and scavenging of free radicals.

Prevention of cytogenetic, histochemical and biochemical alterations in Oreochromis niloticus by dietary supplement of sorbent materials

The current study was conducted to evaluate the ability of Egyptian bentonite (EB) and montmorillonite (EM) for the prevention of genotoxicity, histochemical and biochemical changes induced by aflatoxin B(1) (AFB(1)) using the micronucleus (MN) assay, chromosomal aberrations and DNA fragmentation analysis in Tilapia fish. Six groups of fish were treated for 3 weeks and included the control group, AFB(1)-treated group and the groups treated with EB or EM alone or in combination with AFB(1). At the end of experiment period, blood samples were collected for MN, testosterone and biochemical assays. Chromosomal aberrations were determined in kidney tissues, DNA fragmentation test was determined in liver and testis, whereas histochemical study was carried out on liver, testis and gills. The results indicated that a significant decrease in total protein, albumin, globulin, testosterone and DNA content in liver, gills and testis accompanied with a significant increase in number of micronucleated erythrocytes (MnRBCs), total chromosomal aberrations in kidney and DNA fragmentation in testis and liver of fish received AFB(1) alone. Fish treated with EB or EM alone were comparable to the control regarding the biochemical parameters except testosterone in EB-treated group which was significantly decreased. Both clays did not induce any significant differences in number of MnRBCs, chromosomal aberrations in the kidney, DNA fragmentation in testis, but not in liver of EB-treated group. The combined treatment with AFB(1) and EB or EM succeeded to improve all the tested parameters towards the control values although it did not normalize them. Moreover, the improvement was pronounced in the group received EM plus AFB(1). It could be concluded that EB and EM have the ability to tightly bind AFB(1) in the gastrointestinal tract of fish resulting in decreasing its bioavailability. Moreover, the two tested clays were safe and can be used as potential aflatoxin binders in animal feed.

Effectiveness of activated carbon and Egyptian montmorillonite in the protection against deoxynivalenol-induced cytotoxicity and genotoxicity in rats

This study was conducted to prepare and characterize activated carbon (AC) and to evaluate its protective effect against deoxynivalenol (DON) toxicity in rats compared to Egyptian montmorillonite (EM). AC was prepared using a single-step chemical activation with phosphoric acid (H3PO4). The resulted AC has a high surface area and a high total pore volume. Male Sprague-Dawley rats were divided into 6 groups (n = 10) and treated for 3 weeks as follow: the control group, the groups fed AC or EM-supplemented diet (0.5% w/w), the group treated orally with DON (5 mg/kg b.w.) and the groups fed AC or EM-supplemented diet and treated with DON. Blood and liver samples were collected for different analyses. Treatment with DON increased liver function enzymes, lipid peroxidation, tumor necrosis factor α, DNA fragmentation, decreased hepatic glutathione content, up regulating mRNA Fas and TNF-α genes expression and increased micronucleated polychromatic erythrocytes and normochromatic erythrocytes in bone marrow. Co-treatment of DON plus AC or EM succeeded to normalize the levels of the biochemical parameters, reduced the cytotoxicity of bone marrow and ameliorated the hepatic genotoxicity. Moreover, AC was more effective than EM and has a high affinity to adsorb DON and to reduce its cytotoxicity and genotoxicity.

Inhibition of cadmium- induced genotoxicity and histopathological changes in Nile tilapia fish by Egyptian and Tunisian montmorillonite clay

Cadmium (Cd) is an important inorganic toxicant widely distributed in the environment because of its various industrial uses. The aims of the current study were to investigate the efficacy of purified Egyptian and Tunisian montmorillonite clays (EMC and TMC) to inhibit genotoxicity and histological alterations induced by cadmium chloride (CdCl2) utilizing the Nile tilapia fish as an in vivo model. Chromosomal aberrations (CAs), micronucleus (MN) frequencies and DNA fingerprinting profile were genotoxic end points and histopathological changes that were used in this investigation. Six groups of fish were treated for 2 weeks and included control group, CdCl2-treated group and groups treated with EMC or TMC alone or in combination with CdCl2. The present results revealed that, treatment of fish with CdCl2 exhibited significant increased in the number of micronucleated erythrocytes (MnRBCs), frequency of CAs and instability of genomic DNA. Treatment of EMC and TMC in combination with CdCl2 significantly reduced the frequency of MnRBCs by the percentage of 53.28% and 60.77% and the frequency of CAs by 43.91% and 52.17% respectively. As well as, normalized DNA fingerprinting profile and significantly improved histopathological picture induced by Cadmium treatment. It is worth mention that both clays have the ability to tightly bind CdCl2 and decreased its cytotoxicity and genotoxicity; however, Tunisian clay was more efficient in binding with the CdCl2 than Egyptian clay.

Lactobacillus plantarum alleviate aflatoxins (B 1 and M 1 ) induced disturbances in the intestinal genes expression and DNA fragmentation in mice

&NA; This study aimed to assess the disturbances in intestinal genes expression and DNA fragmentation in mice treated orally with aflatoxin B1 (AFB1) or aflatoxin M1 (AFM1) and the protective activity of Lactobacillus plantarum (LP). Male Balb/c mice were divided into 6 groups including the control group, the group treated with 2 mg/kg b.w of LP (2 × 109 cfu/mL), the groups treated with AFB1 or AFM1 (100 &mgr;g/kg b.w), and the groups treated with AFB1 or AFM1 during, after or before LP. Small intestines were collected for the determination of DNA fragmentation, gene expression and target protein content. The results showed that AFB1 or AFM1 increased DNA fragmentation, down regulated the expressions of caspase‐3, caspase‐9, CYP3A13, Bax and p53 as well as up‐regulated the expression of TNF‐&agr; and Bcl‐2 and their target proteins. LP succeeded to alleviate the disturbances in DNA fragmentation and the expression of these genes. The improvement was more pronounced in the group co‐administered with the toxins plus LP. It could be concluded that AFB1 and AFM1 induced disturbances in intestinal function via the disturbances in DNA fragmentation and genes expression. LP induced a potential protective effect and is considered a promising agent against the genotoxicity induced by these mycotoxins. HighlightsAflatoxins B1 and M1 (AFB1, AFM1) are well known carcinogenic mycotoxins.Both aflatoxins induced severe disturbances in the intestinal function.They induced disturbances in the expression of several genes and their target protein.Lactobacillus plantarum protect against the disturbance induced by these aflatoxins.This protection was pronounced when the bacteria was administrated plus aflatoxins.

Abstract B76: Protective role of Aquilegia vulgaris L extract against fumonisin‐induced oxidative stress and cytotoxicity in rats

Fumonisins (FB) are mycotoxins produced by the fungus Fusarium verticillioides, which commonly infects corn and other crops worldwide. Exposure to FB is known to have toxic and carcinogenic effects in different animal species, and to express toxicity in cells via the induction of oxidative stress. The aim of the current study was to evaluate the protective effects of the ethanol extract of Aquilegia vulgaris L against the oxidative stress and the genotoxicity using the chromosomal aberrations in somatic cells assay and random amplified polymorphism DNA (RAPDPCR) in FBtreated rats. Sixty mature female Sprague‐Dawley were divided into six treatment groups and treated for 4 weeks as follow: the control group, the group fed fumonisin‐contaminated diet (200 mg/kg diet), the groups treated orally with the extract (5 and 10 mg/kg b.w) and the groups fed fumonisin contaminated diet and treated with the extract at the two doses. The results indicated that animals treated with fumonisin alone disturbs lipid profile in serum, increases Sa/So ratio, induces bone marrow cytotoxicity, increases DNA and RNA in liver accompanied with significant changes in histological picture. The extract alone at the two tested doses did not induce any significant changes in the biochemical or histological picture. The combined treatment showed significant improvements in all tested parameters and histological pictures in the liver tissues. Moreover, this improvement was more pronounced in the group received the high dose of the extract. It could be concluded that the ethanol extract of A. vulgaris induces its protective effect via increase the antioxidant capacity and the inhibition of lipid peroxidation. Citation Information: Cancer Prev Res 2010;3(1 Suppl):B76.