[Effects of 2,2', 4,4' -tetrabromodiphenyl ethers on oxidative stress and DNA damage in SH-SY5Y cells].
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To investigate the effects of 2, 2', 4,4 '-polybrominated diphenyl ethers (PBDE-47) on SH-SY5Y cells were oxidative stress and DNA damage in human neuroblastoma cells (SH-SY5Y cells).cultured in DMEM supplemented with 10% fetal bovine serum at 37 degrees C in a humidified incubator with 95% air and 5% CO2. The rate of cellular survivors, LDH leakage, contents of MDA and GSH, activity of SOD, and DNA damage were measured after exponentially growing cells were incubated with 1, 2, 4, 6, 8 and 10 microg/ml PBDE-47 for 24 hours in vitro.The rate of cellular survivors in the low dose PBDE-47-treated groups (l microg/ml and 2 microg/ml) were higher than the control group (P <0.05), but those in the high dose PBDE-47-treated groups (4, 6, 8 and 10 microg/ml) were significantly lower than the control group (P < 0.05). Compared with the control group, the GSH content were significantly decreased (P < 0.05) and DNA tail moment were significantly increased with increasing PBDE-47 concentrations. The MDA content in the high PBDE-treated groups (4, 8 and 10 microg/ml) were notably higher than the control group and increased with increasing PBDE-47 concentrations (P < 0.05). In the high PBDE-treated groups (4, 6, 8 and l0 microg/ml), the LDH leakage were markedly higher and the SOD activity were markedly lower than the control group (P < 0.05). The percentage of DNA in the tail in the high PBDE-treated groups (6,8 and 10 microg/ml) were visibly higher than the control group (P < 0.05).PBDE-47 could induce oxidative stress and DNA damage in SH-SY5Y cells. The oxidative stress may play an important role in the DNA damage induced by PBDE-47.Keywords:
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To explore the protective effects of exogenous reduced glutathione (GSH) on the genotoxicity and oxidative stress induced by sodium arsenite (NaAsO2).Human lung adenocarcinoma A549 cells were divided into several groups according to the treatments as follow: untreated group, single NaAsO2 or GSH treated group and the groups co-treated with NaAsO2 and different concentrations of GSH. Then the differences of cell viability, level of oxidative stress, DNA and chromosomal damage were compared after single or combined treatments.The rate of cell survival and colony formation, the contents of GSH and the activity of SOD in NaAsO2 treated group were significantly lower than the non-treated group, whereas, the level of ROS, comet rate, OTM and the frequency of micronucleus in NaAsO2 treated group were significantly higher than that of control group (P < 0.05). At the 0.5-5 mmol/L concentrations of GSH, these indicators of the co-treatment groups did not show any significant difference when compared with single NaAsO2-treated group. However, at the 10 mmol/L and 20 mmol/L concentrations of GSH, the NaAsO2-induced toxic effects were found to be weakened by GSH, but it was still significantly lower than that in the control group (P < 0.05). Moreover, there was no significant difference among co-treated groups and control group on the cell viability, colony formation, level of oxidative stress and DNA and chromosomal damage at the 40 or 50 mmol/L of GSH.High concentrations of exogenous GSH can significantly decrease the cytotoxicity of NaAsO2, and alleviate DNA and chromosomal damage and the level of oxidative stress.
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Several epidemiological and experimental studies has been reported that lutein (LT) presents antioxidant properties. Aim of the present study was to investigate the protective effects of LT against oxidative stress and DNA damage induced by cisplatin (cDDP) in a human derived liver cell line (HepG2). Cell viability and DNA-damage was monitored by MTT and comet assays. Moreover, different biochemical parameters related to redox status (glutathione, cytochrome-c and intracellular ROS) were also evaluated. A clear DNA-damage was seen with cDDP (1.0μM) treatment. In combination with the carotenoid, reduction of DNA damage was observed after pre- and simultaneous treatment of the cells, but not when the carotenoid was added to the cells after the exposure to cDDP. Exposure of the cells to cDDP also caused significant changes of all biochemical parameters and in co-treatment of the cells with LT, the carotenoid reverted these alterations. The results indicate that cDDP induces pronounced oxidative stress in HepG2 cells that is related to DNA damage and that the supplementation with the antioxidant LT may protect these adverse effects caused by the exposure of the cells to platinum compound, which can be a good predict for chemoprevention.
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Ionizing radiation (lR) has been extensively used as therapy and diagnostic modality to detect abnormalities inside human body, but elicit both beneficial and deleterious role. Interaction between and IR and cells can leads to production of free radicals thus causing oxidative stress. By using animal model system, our aim is to assess molecular DNA damage and oxidative stress in mice intestinal tissue following 50% watermelon juice supplementation for 14 days. Twenty four (24) of6 weeks old male ICR mice were randomly selected into four groups, which are negative control group (-ve), antioxidant group (Aox), radiation group (Rx) and treatment group (Tx). Cx were treated with normal diet and filtered tap water; Aox were treated with normal diet and 50% watermelon juice; Rx were treated with normal diet, filtered tap water and irradiated with 100 IlGy x-ray; Tx were treated with normal diet, 50% watermelon juice and irradiated with 100 IlGy x-ray. After 14 days, the levels of superoxide dismutase (SOD), reduced glutathione (GSH) and malondialdehyde (MDA) in intestinal tissues were elucidated by using biochemical analysis and comet assay for demonstration of oxidative stress and DNA damage. Comet assay revealed significant reduction of DNA damage in 50% watermelon juice supplemented group compared to radiation group (p=0.00) and pair-wise relationship between Aox and Cx showed significant difference with p=0.003. Between Rx and Cx, the DNA damage were statistically significant at p=O.OO. GSH levels for Rx and Tx yield significant reduction compared to Cx with p=O.OO and p=O.OO respectively. Significant reduction of DNA damage confirmed ameliorative effect of 50% watermelon juice while reduction of GSH level suggested a high consumption of natural antioxidant to combat oxidative stress prior to IR exposure. The outcomes highlighted that supplementation of 50% watermelon juice for 14 days have a radioprotective properties against DNA damage induced by IR.
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Abstract DNA strand breaks [as determined by the conventional and formamidopyrimidine glycosylase (FPG)‐modified Comet assay] and antioxidant defense status [as indicated by superoxide dismutase (SOD) activity and reduced glutathione (GSH) concentration] were evaluated in healthy adult chub ( Leuciscus cephalus ) after exhaustive exercise [swimming to their critical swimming speed ( U crit ), twice in succession with a 40 min rest period between] vs. confined (unexercised) control fish. The conventional Comet assay revealed significantly higher DNA strand breaks in all the tissues (blood, liver, and gill), with the highest increase over background evident in the epithelial gill cells of swum fish compared to the controls. Moreover, when the FPG‐modified Comet assay was conducted to reveal specific oxidative lesions, the gill cells of exercised fish sustained the highest level of oxidative DNA damage in comparison to the control. Data on tissue antioxidant defense mechanism were less conclusive, with no significant differences in the tissue levels of SOD or GSH. This suggests that either the degree of oxidative stress was not great enough to evoke a response in terms of defense mechanisms or the timescale of antioxidant defense response was somewhat different from the time between the application of stress and subsequent tissue sampling. From the swimming data, U crit was significantly lower on the second assessment compared to the first (repeat ratio: 0.76), suggesting that the fish were exercised to a level which was not sustainable. Overall, these findings support the theory that acute extreme exercise could result in oxidative stress and associated DNA damage in fish. These observations suggest that fish living in fast flowing and polluted rivers are at increased risk of DNA damage. Environ. Mol. Mutagen., 2006. © 2006 Wiley‐Liss, Inc.
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Triclosan (TCS) is used as an antimicrobial agent and has been widely dispersed and detected in the aquatic environment. However, it remains uncertain whether TCS is genotoxic or not. In this study, the acute toxicity of TCS in goldfish (Carassius auratus) was studied. Then, based on the results for acute toxicity, other goldfish were exposed to various concentrations of TCS (control, DMSO control, and 1/4, 1/2, and 1/8 LC50) for 14 days, and the effects on genetic toxicity were evaluated using micronucleus (MN) and nuclear abnormalities (NA) frequencies in peripheral blood and the comet assay in the liver of the goldfish. In addition, malondialdehyde (MDA), reduced glutathione (GSH), catalase (CAT), and total antioxidant capacity (T-AOC) in the liver were assayed to evaluate oxidative stress and the possible mechanism of genotoxicity. The 96 h median lethal concentration of TCS was 1111.9 µg/l. After 14 days of exposure, the MN and NA frequencies were significantly increased in peripheral blood of the TCS-treated groups compared with the solvent control, and the comet tail moment and MDA in the liver in the highest dose of TCS groups were also significantly high. Meanwhile, an evident change in GSH, CAT, and T-AOC of the liver was found as the TCS exposure concentration increased. The results showed that TCS caused oxidative stress and a genotoxic response in goldfish, suggesting that it presents a potential ecotoxicological risk to aquatic ecosystems.
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Neuropathophysiology research is receiving considerable attention. Studies have demonstrated that under oxidative stress, reactive oxygen species (ROS) generated at high levels inducing cellular and DNA damage, thereby resulting in apoptosis of neuronal cells. This is implicated in the etiology of several neurodegenerative and neurodevelopmental disorders. This study was undertaken to examine the role of glutathione as a Neuroprotective bioactive compound on hydrogen peroxide-induced apoptosis. Assessment of DNA damage with the help of Comet assay (single cell gel electrophoresis) and DNA fragmentation Assay were carried out on cultured SH-SY5Y neuroblastoma cells. The treatment with glutathione markedly attenuated hydrogen peroxide-induced cell viability loss and apoptotic neuronal cell death. These results provide evidence that glutathione may act as a significantly bioactive compound and support the possibility that it may be important in health and disease, and for protection against DNA damage by oxidative stress.
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Sulfur mustard
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Alternariol
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α,β‐Unsaturated carbonyl compounds occur in food and other environmental media. Due to their reactivity with cellular nucleophiles (e.g. Michael adduct formation with DNA bases and with glutathione) they might represent a potential health risk. In this study, induction of oxidative DNA damage was investigated in mammalian cells, as a consequence of glutathione depletion induced by selected food relevant 2‐alkenals, including E‐(2)‐hexenal (HEX), (2E,4E)‐2,4‐hexadienal (HEXDI) and (E)‐2‐cinnamaldehyde (CA) and the cyclic analogue 2‐cyclohexen‐1‐one (CHX). Oxidative DNA breakage was monitored with the Comet assay, using treatment with formamidopyrimidine‐DNA glycosylase (FPG). Total cellular glutathione (tGSH) was determined in a kinetic, photometric assay. After 1 h incubation of V79 cells with HEX (100 µM) and CHX (300 µM), HEXDI and CA (300 µM each), tGSH was depleted down to <20% of control (viability >85%). Under these conditions, FPG‐sensitive sites were not observed; moderate direct DNA breakage, however, was detectable. During 3 h post‐incubation (without test compound) distinct oxidative DNA breakage occurred in HEX‐ and CA‐, but not in CHX‐ and HEXDI‐pretreated cells. Direct DNA breakage was markedly diminished, most probably by repair processes, and tGSH concentrations were observed to increase again within 3 h post‐treatment. The results give strong evidence for alkenal‐mediated oxidative stress contributing to cytotoxic/genotoxic cell damage. The extent of oxidative stress appears to be influenced by structure‐specific properties of the alkenals.
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