Effects of chronic administration of doxorubicin on myocardial beta-adrenergic receptors

The effects of multiple doses of doxorubicin (DXR) on myocardial beta-adrenergic receptor density and dissociation constant were investigated in male Sprague Dawley rats. The rats received DXR (2 mg/kg) or vehicle weekly by the SC route for 13 weeks. One group of DXR-treated rats plus corresponding controls were sacrificed at 14 weeks, one week after the last dose. Another group of DXR-treated rats plus corresponding controls were sacrificed at 19 weeks, six weeks after the last dose. The myocardial beta-adrenergic receptor was characterized by radio-ligand binding studies using [125I]iodocyanopindolol. Beta--receptor densities in DXR-treated rats of 7.0 and 7.4 fm/mg protein were unchanged from control levels of 7.2 fm/mg protein at both 14 and 19 weeks, respectively. Receptor dissociation constants in DXR-treated rats of 36.7 and 36.9 pM were increased over control levels of 24.6 and 30.0 pM at 14 and 19 weeks, respectively. However, the change in dissociation constant is only significant at 14 weeks. The increased dissociation constants suggest diminished agonist binding affinity of the myocardial beta-receptor. This impaired response of the receptor to catecholamines would tend to diminish the ability of myocardium to adequately respond to adrenergic stimuli.     

Serum creatine phosphokinase levels in rats following cold injury

The validity of serum creatine phosphokinase (CPK) as an index of tissue damage due to cold injury was tested in the rat. Immersion of the feet in the freezing mixture (salt and ice, –19°±1°C) for one or five minutes produced reversible and irreversible damage, respectively. The increase in CPK activity over a 5-hr period following freezing showed that the severity and extent of cold injury was associated with the amount of CPK released into the bloodstream. -Exposure of the whole rat to cold air at –29°C for three hours resulted in a highly variable increase in serum CPK two hours after exposure which was related to the degree of hypothermia during the exposure. The extent of hypothermia was also related to the amount of tissue loss estimated two weeks after the exposure. These findings show that the increase in serum CPK after cold injury reflects subsequent tissue loss and might conceivably be useful in early classification of cold injury.

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Zusammenfassung Die Verwendbarkeit der Serumkreatinphosphatase (CPK) als Indikator von Gewebsschäden nach Erfrierungen wurde an der Ratte untersucht. Eintauchen der Pfote in eine Kältemischung (Salz und Eis, –19°±1°C) über Perioden von 1–5 Minuten führte zu reversiblen und irreversiblen Gewebsschädigungen. Der Anstieg der CPK-Aktivität 5 Stunden nach Beginn der Erfrierungen zeigte, dass die Schwere und das Ausmass des Kälteschadens im Zusammenhang stand mit der in das Blut ausgetretenen CPK-Menge. - Aussetzen von Ratten während 3 Stunden bei –29°C führte zu einem Anstieg der Serum CPK 2 Stunden später. Die Werte wiesen eine grosse Streuung auf, die im Zusammenhang stand mit dem Ausmass der Hypothermie während der Belastung. Die Hypothermie war positiv korreliert mit der Menge des Gewebsverlustes 2 Wochen später. Danach ist der Anstieg der Serum-CPK nach Erfrierungen Ausdruck der Gewebsschädigung und kann eventuell zur frühen Beurteilung von Erfrierungen dienen.

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Resume Des rats ont servi de bêtes d'essai pour vérifier si l'analyse de la phosphatase de la créatine du sérum (CPK) est utilisable comme indicateur de dommages des tissus après des phénomènes de gelures. Le fait de placer les pattes des rats dans un mélange réfrigérant (mélange de glace et de sel; température –19° à ± 1°C) durant des périodes de 1 à 5 minutes a conduit à des dégâts réversibles et irréversibles des tissus. L'augmentation de l'activité de CPK cinq heures après le début des gelures a montré que l'importance des lésions était en liaison directe avec les quantités de CPK présentes dans le sang. L'exposition de rats durant 3 heures à une température de –29°C a conduit à une augmentation du taux de CPK du sérum deux heures après. Les valeurs recueillies présentent cependant une très grande variabilité qui est en relation directe avec l'importance de l'hypothermie durant la période de contrainte. Cette hypothermie est en corrélation positive avec la quantité de tissu perdu deux semaines après l'exposition. Il ressort de cette expérience que l'augmentation de CPK dans le sérum sanguin après des gelures peut être utilisée comme expression des dégâts subis par les tissus et éventuellement pour la détermination précoce de phénomènes de gelures.

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DCIEM Research Paper 913     

The effect of thyroxine administration on lipid peroxidation in different tissues of rats with hypothyroidism

Thyroid dysfunctions bring about pathological changes in different organs of the body. Findings obtained from in vivo and in vitro studies point out that thyroid hormones have a strong impact on oxidative stress. The present study was conducted to demonstrate how high-dose thyroxin administration for one week affected oxidative damage formed in experimental hypothyroidism. The study was carried out with 30 Spraque-Dawley species male rats. The experimental animals were divided into 3 groups (Group 1, control; Group 2, hypothyroidism; Group 3, hypothyroidism + thyroxine administration). Malondialdehyde (MDA) and glutathione (GSH) levels were determined in cerebral, hepatic and cardiac tissues after the experimental period. MDA and GSH levels in cerebral, hepatic and cardiac tissues of hypothyroidism + thyroxine supplemented group were higher than those in the control and hypothyroidism groups (p<0.001). The same parameters were higher in the control group than those in the hypothyroidism group (p<0.001). The results of the present study show that hypothyroidism reduced the oxidative damage in cerebral, hepatic and cardiac tissues of rats. However, high-dose thyroxine administration in addition to induced hypothyroidism increased oxidative damage in the same tissues and that this damage could not be prevented despite the increase in the antioxidant system activity.     

Effects of hyperthyroidism induced by L-thyroxin administration on lipid peroxidation in various rat tissues

Thyroid dysfunctions are associated with many pathological signs in the body. One of these is lipid peroxidation that develops due to over- or under-secretion of thyroid hormones. The present study was conducted to determine lipid peroxidation that develops in different tissues including the brain, liver and heart of rats in experimental hyperthyroidism induced by L-thyroxin. The study was carried out on 30 male Sprague-Dawley rats. They were divided into three groups as control, sham hyperthyroidism and hyperthyroidism. Malondialdehyde (MDA) and glutathione (GSH) levels in rat tissues were determined at the end of a 3-weeks period of L-thyroxin administration. It was observed that MDA levels in the hyperthyroidism group were significantly higher in the cerebral cortex, liver and ventriculer tissue of heart (p < 0.001) than in the control and in sham hyperthyroidism groups. GSH levels were higher in the hyperthyroidism group than in control and sham hyperthyroidism groups in all tissues (p < 0.001). Results demonstrate that hyperthyroidism induced by L-thyroxin activates both oxidant and antioxidant systems in cerebral, hepatic and cardiac tissues. However, the increase in antioxidant activity cannot adequately prevent oxidative damage.     

Effects of pyrroline and pyrrolidine nitroxides on lipid peroxidation in heart tissue of rats treated with doxorubicin

Protection from doxorubicin-induced lipid peroxidation in vivo by two pyrroline and pyrrolidine nitroxides, Pirolin, PL, and Pirolid, PD, was examined in the heart tissue of rats treated with this drug. The level of lipid peroxidation was estimated on the basis of MDA content. A considerable (three-fold) increase in the MDA amount was found in heart homogenates from rats injected with doxorubicin, whereas no significant changes in MDA content compared to control were observed in cardiomyocytes treated with the nitroxides (Pirolin or Pirolid) only. Pirolin injected simultaneously with doxorubicin showed antioxidative effect and markedly attenuated lipid peroxidation in the heart tissue caused by this drug. In contrast to Pirolin, structurally related Pirolid was ineffective in the protection of heart myocytes from DOX-induced lipid peroxidation.     

Hesperidin, a flavanone glycoside, on lipid peroxidation and antioxidant status in experimental myocardial ischemic rats

Myocardial infarction continues to be a leading cause of mortality world-wide. Novel therapies are needed to treat the myocardial ischemia. This study was undertaken to evaluate the cardioprotective role of hesperidin on isoproterenol-induced myocardial ischemia in rats. Myocardial ischemia was induced by subcutaneous injection of isoproterenol hydrochloride (85 mg/kg body weight), for two consecutive days. Isoproterenol-administered rats showed elevated levels of cardiac markers (aspartate transaminase, alanine transaminase, lactate dehydrogenase, creatine kinase, creatine kinase-MB, cardiac troponins T and I) when compared with control and hesperidin treatment groups (100, 200 and 400 mg/kg body weight). The serum levels of cardiac markers were significantly reduced at the doses of 200 mg and 400 mg. All further experiments were carried out at the 200 mg dose. Lipid peroxidation markers (thiobarbituric acid reactive substances, lipid hydroperoxides and conjugated dienes) were elevated significantly in the plasma and heart whereas non-enzymic antioxidants (vitamin C, vitamin E and reduced glutathione) were decreased significantly. Activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase declined significantly in the heart of ischemic rats. However, after hesperidin treatment, all the above parameters reverted to normal levels. This study demonstrated that the cardioprotective effect of hesperidin on ischemic rats could be due to its anti-lipid peroxidative and antioxidant properties.     

Effect of chronic fluorosis on lipid peroxidation and histology of kidney tissues in first- and second-generation rats

This experiment was designed to investigate the lipid peroxidation and histological effects of chronic fluorosis on first- and second-generation rat kidney tissues. Sixteen virgin female Wistar rats were mated with eight males (2∶1) for approx 12 h to obtain first-generation rats. Mating was confirmed by the presence of sperm in vaginal smears. Sperm in vaginal smears was observed in 10 of 16 rats (d 0). These rats were identified as pregnant and included in this experiment. Pregnant rats were divided into two experimental groups (control and fluoride-supplemented), each containing five rats. The pregnant rats in the fluoride-supplemented group were exposed to 30 mg/L sodium fluoride (NaF) in commercial drinking water containing 0.07 mg/L NaF throughout the gestation and the lactation periods. After the lactation period, young animals (first generation [F1]) were exposed to the same amount of NaF in drinking water for 4 mo. At the end of the 4-mo experimental period, nine randomly chosen male rats (F1) were sacrificed, and the kidneys were removed for the histological and lipid peroxidation examinations. The remaining eight female rats were mated with four males (2∶1) for approx 12 h to obtain second-generation rats. Six female were identified as pregnant, and treated similarly throughout the gestation and the lactation periods. After the lactation period, the young male rats (second-generation male rats [F2]) were also treated similarly for 4 mo. At the end of the 4-mo experimental period, nine randomly chosen male rats (F2) were sacrificed, and the kidneys were removed for the histological and lipid peroxidation examinations. The rats in the control groups underwent the same procedure without NaF supplementation. It was found that the plasma fluoride and kidney TBARS levels of fluoride-supplemented F1 and F2 rats were higher than controls. Hydropic epithelial cell degenerations and moderate tubular dilatation were observed in some proximal and distal tubules. There were markedly focal mononuclear cell infiltrations and hemorrhage at some areas of the interstitium, especially at the corticomedullar junction. Mononuclear cell infiltrations were also evident in some peritubular and perivascular areas. Most of the vascular structures were congestive. Many Bowman capsules were narrowed. The severe degenerative changes in most of the shrunken glomerules and vascular congestion were also observed. It is concluded that chronic fluorosis causes a marked destruction in kidney tissues of F1 and F2 rats by causing lipid peroxidation. Department of Orthopedics, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey     

Short-term thyroxine administration leads to lipid peroxidation in renal and testicular tissues of rats with hypothyroidism

Thyroid dysfunction brings about pathological changes in different organs of the body. The aim of the present study was to examine how experimental hypothyroidism and additional short-term high-dose thyroxine administration (one-week) affected lipid peroxidation in renal and testicular tissues of rats. The study was carried out on 30 male Spraque-Dawley rats. The experimental animals were divided into 3 groups as control, hypothyroidism and hypothyroidism + thyroxine administration. Both malondialdehyde (MDA) and glutathione (GSH) levels were lower in renal and testicular tissues of the hypothyroidism group than the control and hypothyroidism + thyroxine administration groups and the levels in hypothyroidism + thyroxine administration group were higher than those in the control and hypothyroidism groups (p < 0.001). Results of the study demonstrate that hypothyroidism reduced oxidant stress in kidney and testis tissues, but short-term, high-dose thyroxine administration in addition to hypothyroidism increased oxidant stress in the same tissues of rats.     

厚壳贻贝抗氧化酶活性和脂质过氧化物含量对Cd和Aroclor 1254胁迫的响应

以厚壳贻贝(Mytilus coruscus)为目标生物,研究了不同浓度Cd2+(0.194、0.388、0.775 mg·L-1)、Aroclor 1254(0.005、0.01、0.05 mg·L-1)单一和复合胁迫对其消化盲囊抗氧化酶活性及脂质过氧化产物丙二醛(MDA)含量的影响。结果表明:(1)单一和复合胁迫均可导致厚壳贻贝抗氧化酶的活性先升高后抑制。高浓度组(0.775 mg·L-1Cd2+、0.05mg·L-1Aroclor 1254)在单一胁迫第1天时,抗氧化酶的活性即达峰值。抗氧化酶活性的下降幅度和速率与胁迫物质作用时间和浓度呈正相关。(2)SOD、GSH-Px对胁迫的敏感性高于CAT。(3)各胁迫处理组MDA含量随胁迫时间延长均显著升高(P0.01)。单一胁迫的效应与剂量和时间呈正相关。(4)复合胁迫的效应强于单一胁迫。推测高于环境水平的Cd2+、Aroclor 1254对厚壳贻贝可产生明显氧化胁迫,抗氧化防御系统可以作为海洋环境重金属-有机物复合污染监测的潜在生物标志物。     

Protective effects of thymol on altered plasma lipid peroxidation and nonenzymic antioxidants in isoproterenol-induced myocardial infarcted rats

This study evaluates the protective effects of thymol on altered plasma lipid peroxidation products and nonenzymic antioxidants in isoproterenol (ISO)‐induced myocardial infarcted rats. Male albino Wistar rats were pre and cotreated with thymol (7.5 mg/kg body weight) daily for 7 days. ISO (100 mg/kg body weight) was subcutaneously injected into rats on 6th and 7th day to induce myocardial infarction (MI). Increased activity/levels of serum creatine kinase‐MB (CK‐MB), plasma thiobarbituric acid reactive substances, lipid hydroperoxides, and conjugated dienes with decreased levels of plasma reduced glutathione (GSH), vitamin C, and vitamin E were observed in ISO‐induced myocardial infarcted rats. Pre and cotreatment with thymol (7.5 mg/kg body weight) showed normalized activity of serum CK‐MB and near normalized levels of plasma lipid peroxidation products, reduced GSH, vitamin C, and vitamin E in myocardial infarcted rats. Furthermore, the in vitro study on reducing power of thymol confirmed its potent antioxidant action. Thus, thymol protects ISO‐induced MI in rats by its antilipid peroxidation and antioxidant properties. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:368–373, 2012; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21431     

Effects of combined irradiation and doxorubicin treatment on cardiac function and antioxidant defenses in the rat

Combined radiotherapy and chemotherapy have represented a major advance in the therapeutic management of cancer therapy. However, the combination of doxorubicin (DXR) and cardiac irradiation (IRR) could precipitate the unexpected expression of congestive heart failure. Oxidative lesions induced by IRR and DXR could represent one of the pathogenic factors of myocardial dysfunction. Our investigations were performed to evaluate in the rat: 1) cardiac functional changes, 2) cardiac and plasma peroxidative damage and antioxidant defenses variations, that occur 24 h (acute effects) and 30 d (middle term effects) following DXR treatment 1 mg/kg(-1)/day(-1) IP for 10 d and a 1 x 20 Gy cardiac gamma-irradiation. Our results showed that DXR affected heart reactivity as early as the end of its administration, although irradiation exerted no detectable effect. Antioxidant defenses disturbances in hearts of DXR treated rats were characterized by vitamins C and E decreases, catalase activity induction and an increase in lipid peroxidation. Moreover, plasma vitamin C consumption and the lower level of plasma lipid peroxidation attested to the efficient solicitation of antioxidant defenses that probably contributed to the preservation of cardiac function at 24 h. After 30 d, cardiac dysfunction became symptomatic at rest, resulting from DXR cardiac toxicity. In spite of the persistent activation of cardiac catalase activity, antioxidant deficiency and increased plasma and cardiac lipid peroxidation highlighted defenses overtaken. Thus, different physiopathological mechanisms are involved in heart disturbance at acute and middle terms, IRR and DXR acting on distinct targets without disclosing synergistic effects. After 30 d, cardiac and plasma biochemical abnormalities were emphasized by the combined DXR+IRR therapy, pointing out the severity of the damage. Oxidative damage to the heart induced both by irradiation and DXR, may be one of the pathogenic factors of myocardial dysfunction. There is the possibility that the deleterious effects might be limited by the use of pharmacologic antioxidant agents.     

Plasma lipid peroxidation and antioxidant levels in patients with rheumatoid arthritis

In recent years, a great number of studies have investigated the possible role of reactive oxygen species in the aetiology and pathogenesis of rheumatoid arthritis (RA). The aim of this study was to investigate plasma concentrations of vitamin E, beta-carotene, activities of glutathione peroxidase (GSH-Px) and catalase, levels of lipid peroxidation (MDA) and reduced glutathione (GSH) in 36 patients with rheumatoid arthritis (RA) and 22 healthy age-matched controls. The plasma activity of GSH-Px and catalase (p < 0.001), levels of GSH (p < 0.01), concentration of beta-carotene (p < 0.05) and vitamin E (p < 0.001), haemoglobin and hematocrit (p < 0.05) were significantly lower in patients with RA than in controls. The MDA levels (p < 0.01), C reactive protein, rheumatoid factor, anti-streptolysin-o values (p < 0.001), platelet count (p < 0.05) and erythrocyte sedimentation rate (p < 0.001) were higher in the patient group than in the control group. These results provide some evidence for a potential role of increased lipid peroxidation and decreased enzymic and non-enzymic antioxidants in RA by its inflammatory character. These results suggested that oxidant stress plays a very important role in the pathogenesis of RA.     

The effect of lipoic acid on antioxidant status and lipid peroxidation in rats exposed to chronic restraint stress

This study was designed to investigate effect of alpha-lipoic acid (LA) on lipid peroxidation, nitric oxide production and antioxidant systems in rats exposed to chronic restraint stress. Twenty four male Wistar rats, aged three months, were divided into four groups: control (C), the group treated with LA (L), the group exposed to restraint stress (S) and the group exposed to stress and treated with LA (LS). Restraint stress was applied for 21 days (1 h/day) and LA (100 mg/kg/day) was injected intraperitonally to the L and LS groups for the same period. Restraint stress significantly decreased brain copper/zinc superoxide dismutase (Cu,Zn-SOD) and brain and retina glutathione peroxidase (GSH-Px) and catalase (CAT) activities compared with the control group. Thiobarbituric acid reactive substances (TBARS), nitrite and nitrate levels were significantly increased in the tissues of the S group compared with the C group. LA produced a significant decrease in brain and retina TBARS, nitrite and nitrate levels of the L and LS groups compared to their corresponding control groups. LA increased all enzyme activities in the tissues of the LS group compared to the S group. Our study indicated that LA is an ideal antioxidant candidate for the prevention of stress-induced lipid peroxidation.     

Effects of metal ions on the antioxidant enzyme activities, protein contents and lipid peroxidation of carp tissues

1. Studies were performed regarding the effects of CuSO4 in concentrations of 5, 10, 25 and 50 ppm and ZnSO4 in concentrations of 10 and 100 ppm on the antioxidant enzyme activities, lipid peroxidation and protein contents of tissues of common carp (Cyprinus carpio morpha L.) exposed to these pollutants for 24 hr. 2. The results demonstrated that CuSO4 was more toxic than ZnSO4 and that both treatments brought about significant changes in these parameters in carp hepatopancreas (liver), gill and white muscle. 3. An increase of the CuSO4 concentration led to significant decreases in the antioxidant enzyme activities, except that of glutathione peroxidase, which was increased significantly, and significant increases in the lipid peroxidation and protein contents. 4. An increase of the ZnSO4 concentration led to slight changes in the antioxidant enzyme activities, lipid peroxidation and protein contents of carp tissues.     

Effect of seminal plasma antioxidant on lipid peroxidation in spermatozoa, mitochondria and microsomes

Seminal plasma antioxidant inhibited ascorbate/iron-induced lipid peroxidation in spermatozoa, brain and liver mitochondria. The concentration required to produce inhibition in brain and liver mitochondria was high. Denaturation of spermatozoa resulted in complete loss of antioxidant action. Maintenance of native structure was essential for action of seminal plasma antioxidant in spermatozoal lipid peroxidation. The antioxidant inhibited NADPH, Fe3+-ADP induced lipid peroxidation in microsomes and consequences of lipid peroxidation such as glucose-6-phosphatase inactivation were prevented by presence of antioxidant. It did not inhibit microsomal lipid peroxidation induced by ascorbate and iron and xanthine-xanthine oxidase.     

Lipid levels and lipid peroxidation in frog tissues during hyperthermia and hibernation

The content of cholesterol, phospholipids, free fatty acids in the blood, ketone bodies and lipid peroxidation in the liver, brain, myocardium, skeletal muscles of Rana ridibunda were studied. Changes in the lipid content in blood during artificial hypothermia differ from those during hibernation and arousal. The utilization of reserve fats during hibernation is limited, but significantly rises during arousal. The ability to spontaneous lipid peroxidation under conditions of homogenate incubation at 37 degrees C does not increase in the studied tissues during hibernation.     

Chronic methylphenidate administration alters antioxidant defenses and butyrylcholinesterase activity in blood of juvenile rats

Methylphenidate (MPH), a psychostimulant that affects both dopaminergic and noradrenergic systems, is one of the most frequently prescribed treatments for attention-deficit hyperactivity disorder. The present study investigated the effects of chronic administration of MPH on some parameters of oxidative stress, as well as on butyrylcholinesterase (BuChE) activity in blood of young rats. Rats received intraperitoneal injections of MPH (2.0 mg/kg) once a day, from the 15th to the 45th day of age or an equivalent volume of 0.9% saline solution (controls). Two hours after the last injection, animals were euthanized, and blood was collected. Results demonstrated that MPH did not alter the dichlorofluorescein formed, decreased both thiobarbituric acid reactive substances and total non-enzymatic radical-trapping antioxidant, and increased superoxide dismutase and catalase activities, suggesting that this psychostimulant may alter antioxidant defenses. BuChE activity was increased in blood of juvenile rats subjected to chronic MPH administration. These findings suggest that MPH may promote peripheral oxidative adaptations and cholinergic changes.     

Effects of X-ray radiation on lipid peroxidation and antioxidant systems in rabbits treated with antioxidant compounds

The aim of this study was to investigate the effects of supplemental antioxidant vitamins and minerals on lipid peroxidation and on the antioxidant systems in rabbits exposed to X-rays. The rabbits were divided into two experimental groups and one control group, each group containing seven rabbits. The first group (VG) received daily oral doses of vitamin E (460 mg/kg live weight) and vitamin C (100 mg/kg live weight). The second group (MG) was fed a mineral-enriched diet that contained 60 mg manganese chloride, 40 mg zinc sulfate, and 5 mg copper sulfate per kilogram of feed. The third group served as controls and received only a standard diet. Blood samples were obtained before and after the supplementation with vitamins or minerals, as well as before and after irradiation with a total dose of 550-rad X-rays. The blood samples were analyzed for their content of malondialdehyde (MDA), plasma vitamins C and E, retinol, reduced glutathione (GSH), and glutathione peroxidase activity (GPx). After irradiation, the control group showed increased levels of MDA and activity of GPx (p<0.05), whereas the levels of GSH, vitamin C, and vitamin E were decreased. In the VG, the concentration of MDA was lower (p<0.05), and the concentration of GSH and vitamins C and E were higher (p<0.05) when compared to controls. In the MG, the concentrations of MDA, GSH, vitamin C, and retinol were not affected by the mineral administration and radiation. The level of vitamin E in the MG increased with mineral administration (p<0.05), but decreased after irradiation (p<0.05). For the control group, the level of GSH was higher than in the two experimental groups. After irradiation, the VG animals had vitamin E and C levels that were higher than in MG and control groups (p<0.05). The activity of GPx was not affected by vitamin or mineral supplementation or by irradiation. We conclude that the supplementation with antioxidant vitamins and minerals may serve to reinforce the antioxidant systems, thus having a protective effect against cell damage by X-rays.