Vitamin C prevents hyperbaric oxygen-induced growth retardation and lipid peroxidation and attenuates the oxidation-induced up-regulation of glutathione in guinea pigs

Hyperbaric oxygen therapy is used to treat various clinical conditions, but it also causes oxidative damage. The objectives of this study are to determine if increased vitamin C intake can prevent hyperbaric oxygen-induced damage and to determine interactions among vitamin C, glutathione and vitamin E in response to oxidative stress. The growth rates of unexposed guinea pigs fed 1.25 mg vitamin C/day were indistinguishable from that of guinea pigs fed 50 mg vitamin C/day. In contrast, hyperbaric oxygen exposure resulted in growth retardation in guinea pigs fed 1.25 mg vitamin C/day, but it had little effect on the growth rates of guinea pigs fed 50 mg vitamin C/day. Increased vitamin C intake also prevented hyperbaric oxygen-induced lipid peroxidation in the liver. In guinea pigs not exposed to hyperbaric oxygen, levels of vitamin C in tissues were closely related to vitamin C intake, but tissue levels of glutathione and vitamin E were not related to vitamin C intake. However, interactions between vitamin C and glutathione were observed upon chronic hyperbaric oxygen exposure. Chronic hyperbaric oxygen exposure resulted in >2-fold increases in the levels of glutathione in liver and lung of guinea pigs fed 1.25 mg vitamin C/day. In comparison, the oxidation-induced increases in glutathione were significantly attenuated in guinea pigs fed 50 mg vitamin C/day. These data show that increased intake of vitamin C can prevent or alleviate the hyperbaric oxygen-induced damage. The interactions between vitamin C and glutathione upon hyperbaric oxygen exposure indicate that there is a homeostatic regulation of antioxidant capacity in guinea pig tissues.     

Clinico-Pathologic Findings in Young Pigs Fed Different Levels of Selenium,Vitamin E and Antioxidant

A randomized, blocked 23 factorial experiment was conducted with 48 young pigs. The treatment factors were: 2 levels of selenium (55 and 115 µg/kg), 2 levels of vitamin E (3 and 53 mg/kg) and 2 levels of the antioxidant feed additive Ethoxyquin (0 and 150 mg/kg). All pigs were kept in single pens and fed ad libitum throughout the experimental period of 9 weeks, i.e. from 3 to 12 weeks of age. Plasma, heart, liver and muscle Se levels as well as whole blood glutathione peroxidase activity (EC 1.11.1.9 GSH-Px) were significantly higher in pigs given a dietary supplement of Se than in pigs given no supplement of Se (P ≤ 0.001). The Se-supplemented pigs showed a tendency to lower mean serum transaminase activity (ASAT and ALAT) than unsupplemented pigs, but the influence was significant (P ≤ 0.05) only for the ALAT activity. Blood vit. E levels were higher for pigs receiving a supplement of vit. E than for unsupplemented pigs (P ≤ 0.001), and so was the resistance of red blood cells against lipid peroxidation (ELP), as expressed by lower ELP values. There were no effects of Ethoxyquin supplementation on the biochemical variables included in the study. The histological examination of heart muscle showed that the score for changes was negatively influenced by both Se and vit. E supplement (P ≤ 0.001) and to some extent also by Ethoxyquin supplement (P ≤ 0.05). The histological picture of m. long dorsi was influenced only by the vit. E supplement (P ≤ 0.01). No histological changes were found in the liver in this study. There were inverse relationships between whole blood GSH-Px defluorescence time and blood Se, and between ELP and whole blood vit. E (P ≤ 0.001).     

Effect of vitamins C and E on toxicity and mutagenicity of hexavalent chromium in rat and guinea pig

The effect of simultaneous pretreatment with vitamins C and E on the toxicity and mutagenicity of K₂Cr₂O₇ in rats and guinea pigs was evaluated. Dietary pretreatment of Cr(VI)-intoxicated rats with ascorbic acid or α-tocopherol normalized vitamin C levels in lungs but not in kidneys. The synergistic preventive effect of both vitamins was confirmed in the production of lipoperoxides in Cr(VI)-intoxicated rats. Simultaneous administration of vitamins C and E in guinea pigs prevented the decrease of vitamin C levels provoked by the oxidative effects of Cr(VI). The results of the micronucleus test in bone marrow showed that it was vitamin C that caused the antimutagenic effect against bichromate, in both rats and guinea pigs. The effect of vitamin E was demonstrated only in an increase of the ratio of NCE to PCE, i.e., in a decrease of the cytotoxic but not the mutagenic effects of hexavalent chromium.     

Influence of Diet on Experimental Swine Dysentery. 1. Effects of A Vitamin E and Selenium Deficient Diet Supplemented with 6.8 % Cod Liver Oil

Sixteen growing pigs were fed a vitamin E and selenium deficient diet; half of the animals (Group 2) were given a daily supply of vitamin E and selenium. After having been fed these diets for 53 days, the pigs were infected orally with minced colonic material from cases with typical swine dysentery. This exposure resulted in outbreaks of swine dysentery in both groups. The incubation times were, however, distinctly shorter and the clinical symptoms much more pronounced in Group 1 than in Group 2. The patho^morphological lesions in the colon also differed between the 2 groups. In the pigs of Group 1 evident pseudomembraneous lesions were observed in the spiral colon. In Group 2, the colonic alterations consisted predominantly of a catarrhal enteritis; pseudomembranes occurred in a minor part of colon in only 4 pigs. Both the clinical and the chemical observations and the pathological findings indicated a much better vitamin E and selenium balance in the pigs of Group 2. It is concluded that the treatment with vitamin E and selenium in Group 2 greatly increased resistance to swine dysentery.     

Effect of Dietary Fat Quality and Vitamin E on the Antioxidant Potential of Pigs

A randomized blocked factorial experiment was conducted with 90 young pigs. From 4 to 11 weeks of age the pigs were kept in individual pens and fed a selenium supplemented basal diet consisting mostly of propionic acid treated barley, soybean meal and dried skim milk, and containing < 0.5 mg vitamin E per kg. The treatment factors during this period were 3 dietary levels of added vitamin E (nil, 10 and 30 mg/kg) and a 6 % supplement of fresh or oxidized fat (2/3 lard and 1/3 herring oil). From 11 weeks of age until slaughter at 90 kg the pigs received the vitamin E supplements but no fat or dried skim milk. The basal diet for this later experimental period was based on untreated dry barley. Blood samples collected during the period of investigation were examined for vitamin E and for resistance against erythrocyte lipid peroxidation (ELP) in order to evaluate the antioxidant status. Analysis of variance and Student’s t-test on least squares means showed the ELP to be influenced independently by the vitamin E supplement and, during the fat feeding period, by the quality of the fat supplement, with the highest peroxidation resistance (low ELP) in the groups fed fresh fat and a high level of vitamin E. Blood vitamin E level was only influenced — positively — by the vitamin E supplement although variations in the feed vitamin E level below 10–15 mg vitamin E per kg did not result in corresponding variations in measurable blood vitamin E concentrations. In the same low range of vitamin E in the feed there was a statistically significant difference in ELP values between the different vitamin E treatment groups. No clinical manifestations of selenium-vitamin E deficiency were observed in the pigs. The ELP and the plasma vitamin E levels observed would seem to suggest that a total of 15 mg vitamin E per kg barley-based feed will not always be sufficient for growing pigs.     

Glutathione Peroxidase Activity and Erythrocyte Lipid Peroxidation as Indices of Selenium and Vitamin E Status in Young Pigs

A randomized, blocked 23 factorial experiment was conducted with 48 pigs from sows fed a diet low in selenium and vitamin E. From 3 to 12 weeks of age the piglets were kept in single pens and fed a basic diet consisting mostly of barley, dried skim milk, soybean meal and dried yeast, and containing 55 µg selenium and 3 mg vitamin E per kg. The treatment factors — i.e. feed supplements — were 2 levels of Se (nil, 60 µg/kg), 2 levels of vitamin E (nil, 50 mg/kg), and 2 levels of the feed antioxidant ethoxyquin (nil, 150 mg/kg). Blood samples, collected at termination of the experiment, were examined for glutathione peroxidase activity (GSH-Px) and resistance against erythrocyte lipid peroxidation (ELP) to evaluate Se and vitamin E status, respectively. Analysis of variance showed the GSH-Px activity to be litter-dependent (P < 0.001) and influenced by selenium supplementation (P < 0.001) but not by the other supplements or by interactions between supplements. Resistance against ELP was influenced only by vitamin E supplementation (P < 0.001). GSH-Px and ELP thus seem to be valuable and simple methods for evaluating, respectively, Se status and vitamin E status in growing pigs.     

Effect of Dietary and Injectable Vitamin E on the Antioxidant Status of Pigs

An experiment concerning 6 different vitamin E treatments was conducted with 30 young pigs. From 4 to 15 weeks of age the pigs were kept in individual pens and fed a selenium supplemented basal diet consisting mostly of propionic acid treated barley and soybean meal, and containing 4.4 mg vitamin E per kg. The treatments were periods with or without vitamin E supplement (20 mg/kg) or a vitamin E injection (200 mg). Blood samples collected during the period of investigation were examined for vitamin E and for resistance against erythrocyte lipid peroxidation (ELP) in order to evaluate the antioxidant status. Analysis of variance showed a litter effect on ELP values at all the weekly investigations and a treatment effect from two weeks after the experiment had started. Also the blood vitamin E level was litter dependent and influenced by treatment. Paired comparisons by Student’s t-test showed a delay of 1 to 2 weeks in the effect on ELP of a dietary vitamin E supplement. In contrast, both ELP and vitamin E changed very rapidly (hours) after vitamin E injections. Independently of the vitamin E treatments there was a rise in ELP within the first 2 or 3 weeks after weaning; this was taken as an index of a reduced antioxidant status during that period.     

Effect of vitamin E- and selenium-deficiency on rat liver chemiluminescence.

The role of vitamin E and selenium as protective agents against oxidative stress was evaluated by measuring liver chemiluminescence in situ. Weanling rats fed a vitamin E- and selenium-deficient diet showed liver chemiluminescence that was increased 60 and 100% over control values at 16 and 18 days respectively after weaning. At day 21, the double deficiency led to hepatic necrosis, as observed by optical and electron microscopy, and increased serum levels of lactate dehydrogenase and alanine aminotransferase. Single deficiencies, in either vitamin E or selenium, did not produce liver necrosis but increased liver chemiluminescence. Vitamin E deficiency led to a 23 and 50% increase in liver emission at days 18 and 20 respectively; selenium deficiency produced a 64% increase at day 16. The activity of liver selenium-glutathione peroxidase diminished to 13% of the control value in the rats fed doubly deficient and selenium-deficient diets. Activities of superoxide dismutase, catalase and non-selenium-glutathione peroxidase were not modified by the different diets. These results suggest that oxy-radical generation may play a major role in hepatic necrosis in vitamin E- and selenium-deficiency.     

Relationship between Selenium Content of Forage,Blood and Organs of Sheep,and Lamb Mortality Rate)

The investigation comprised 12 herds, totalling 660 sheep, in the counties of Stockholm and Uppsala. The selenium content of specimens of forage, whole blood, muscle, liver, and kidney was determined by a fluorimetric method. Data on the number of born and dead lambs were collected 4—5 months after lambing. Dead and sick lambs were subjected to necropsy and to clinico-chemical analyses, respectively. The selenium content of hay and corn was lower than 60 ng per g, which some authors regard as the minimum level of Se requirement for protection against white muscle disease in sheep. The analyses of forage mixtures containing oil concentrates and beet pulp showed selenium levels that were about 2—5 times as high as this minimum level. In herds in which the animals had not been treated with selenium preparations prior to sampling, there was a significant positive correlation between the selenium content of the forage and that of whole blood. A relationship was also noted between the selenium content of forage and of organs. In 9 out of the 12 herds the selenium concentrations in whole blood were on the level that is considered to be representative of white muscle disease. A high mortality rate was recorded for lambs whose mothers had low selenium levels in their blood. Prophylactic and curative treatment with selenium plus vitamin E orally and/or parenterally probably reduced lamb mortality rate in the herds fed a low-selenium diet.     

The effect of selenium and/or vitamin E treatments on radiation-induced intestinal injury in rats

Cytotoxic effects of ionizing radiation on gastrointestinal epithelium may be related to oxidative stress. In this study, we wanted to investigate the effects of selenium, vitamin E and selenium plus vitamin E pretreatments prior to whole abdominal irradiation on intestinal injury. Irradiation caused increased lipid peroxide and decreased GSH levels in the intestine. Intestinal superoxide dismutase and glutathione peroxidase activities were increased, but glutathione transferase activity decreased following irradiation. Selenium and/or vitamin E pretreatments ameliorated these disturbances in prooxidant-antioxidant balance. This amelioriation has been verified with histopathological findings. These results indicate that antioxidant pretreatments prior to irradiation may have some beneficial effects against irradiation-induced intestinal injury.     

Effects of Fermentation Conditions and Aging Temperature on Volatile Ester Contents in Wine

In these present experiments, rats were fed a low casein, vitamin E and selenium deficient diet and were killed at various time intervals up to the occurrence of massive liver necrosis. Liver malondialdehyde, and liver and serum vitamin E levels were analyzed simultaneously with a pathological investigation of the features of the liver and measurement of serum GOT and GPT activities. These investigations were also performed on control rats given a high casein, vitamin E defincient diet or a low casein, vitamin E supplemented diet.

The results show that the liver malondialdehyde level was not increased concomitant with the decrease in liver and serum vitamin E level nor with the occurrence of massive liver necrosis. Liver cells of the rats fed on the low casein, vitamin E deficient diet showed swelling of the cytoplasm at the initial stage, and progressive centrilobular lipid deposition was observed by Sudan III stain.     

The influence of dietary selenium and vitamin E on glutathione peroxidase and glutathione in the rat

The effect of dietary selenium (Se) and vitamin E supplementation on tissue reduced glutathione (GSH) and glutathione peroxidase activity has been studied in the rat. Increasing Se intake by 0.4 ppm gave significantly higher enzyme levels in all tissues studied, an effect not influenced by vitamin E intake. Further increasing Se to 4 ppm gave higher enzyme levels in red blood cells only, while in liver was there was a significant decrease in enzyme activity probably reflecting Se hepatotoxicity. In the absence of Se supplements increasing dietary vitamin E to 100 mg/kg diet significantly increased enzyme activity but this effect was modified by simultaneous Se supplementation.Se intake had no effect on GSH levels. Rats on high vitamin E intake 500 mg/kg had a significantly higher tissue GSH level. Dietary Se had a sparing effect on vitamin E, rats supplemented with Se having significantly raised plasma vitamin E levels.These results confirm the role of selenium in glutathione peroxidase and also show that vitamin E influences the activity of the enzyme.     

Effects of Selenium Administration on Erythrocyte and Blood Plasma Glutathione Peroxidase Activity in Goats

The activity of glutathione peroxidase (GSH-Px, E.G. 1.11.1.9.) was determined in heparinized whole blood, blood plasma and washed erythrocytes from goats before and up to 4 weeks after the administration of selenium (0.4 mg/10 kg BW) and vitamin E (20 mg/10 kg BW) or only vit. E (20 mg/10 kg BW). It was found that Se administration caused a significant increase in enzyme activity in whole blood and washed erythrocytes first detected 2 weeks after the intramuscular injection of Se. No changes were observed in plasma from the treated animals. Minor and insignificant changes were seen in the vit. E treated control animals. It is concluded that GSH-Px activity in blood plasma or serum is of no value as a short-term indicator of the selenium status of goats but whole blood is a good indicator of the long-term status.     

Influence of immunomodulation on the development of Listeria monocytogenes infection in aged guinea pigs

We investigated the impact of immunomodulation on the development of listeriosis within an aged population of guinea pigs after an intragastric challenge with Listeria monocytogenes. Supplementation with vitamin E for 35 days significantly increased the level of cytotoxic T cells (CD8(+)), while treatment with cyclosporin A resulted in a 25% decrease of CD8(+) T cells. In the animals receiving the low dose (10(2) CFU) of L. monocytogenes, 50% of the control-group animals became infected. Only 22% of animals receiving the orthomolecular dose of vitamin E became infected, whereas animals that were immunosuppressed had an infection rate of 89%. In the immunosuppressed group three animals (16%) developed listerial infection with a quantifiable bacterial level of 0.3-3 log CFU g(-1) of organ in the spleen and liver. In the high-dose study, the population of L. monocytogenes was consistently 1 log CFU g(-1) lower in the spleen or liver of the vitamin E-supplemented group, compared with the control and cyclosporin A-treated animals. At day 4, a significant increase in the levels of CD8(+) during listerial infection occurred in vitamin E-supplemented animals, suggesting an increased ability to produce CD8(+) T cells. The results suggest that immunomodulation of the host can influence listerial infection within an aged population of guinea pigs.     

Maternal antioxidant concentrations after uncomplicated pregnancies

Background: To analyse the post-partum concentrations of intra- and extra-cellular blood antioxidants in women with uncomplicated pregnancies.

Methods: Whole blood and plasma thiols, plasma vitamin E and C, serum cholesterol and triglyceride, ferric reducing ability of plasma (FRAP) concentrations were compared between women delivered by caesarean section (n=17) or spontaneous delivery (n=10). A repeated mixed model was used for statistical analysis.

Results: The majority of whole blood thiols increased significantly in both groups the first days post-partum. However, within the caesarean group free cysteine, oxidised cysteine, homocysteine and glutathione and plasma cysteine and homocysteine levels dropped significantly after 24 h, while FRAP levels peaked significantly in this group. Plasma vitamin E levels decreased significantly in both groups within 24 to 48 h after delivery. Independent of the way of delivery whole blood and plasma thiols were significantly increased and vitamin E levels were significantly decreased 3 months post-partum while plasma vitamin C levels and FRAP were unchanged compared to ante-partum levels.

Discussion: Decreased plasma vitamin E levels shortly post-partum are associated with decreased lipid peroxidation. The 24 h post-partum drop of some plasma and whole blood thiols in the caesarean group may be due to prolonged fasting.     

Serum Glutathione Peroxidase Activity and Blood Selenium in Pigs

Blood serum glutathione peroxidase activity and blood selenium concentration were measured in blood samples from pigs subjected to experimentally induced selenium deficiency and dietary selenium supplementation on graded levels. A highly significant correlation between blood selenium and serum GSH-Px activity in pigs, especially in selenium deficient pigs, was demonstrated. There was also a strong relationship between blood selenium concentration and serum GSH-Px activity in pigs receiving dietary selenium at graded levels. Serum GSH-Px activity exhibited an excellent close-response relationship to dietary selenium. Linear regression analysis showed that the increased serum GSH-Px activity was a function of the dietary selenium concentration. The fitness of serum in monitoring slight changes of the selenium status of pigs with help of the estimation of GSH-Px activity was discussed. The measurement of serum GSH-Px activity seems to provide a useful and rapid means for defining selenium requirements and for identifying selenium deficiency in growing pigs.     

Effects of intraperitoneally‐administered vitamin E and selenium on the blood biochemical and haematological parameters in rats

The aim of this work was to determine the role of intraperitoneally‐administered vitamin E and selenium on the biochemical and haematological parameters in the blood of rats. Thirty‐two adult male Wistar rats were used in this study. All rats were randomly divided into four groups. The first group was used as the control. The second group was intraperitoneally administered with vitamin E (±‐α‐tocopheroryl acetate, 10 mg day⁻¹), the third group with Se (Na₂SeO₃ 0·2 mg over a day), and the fourth group with vitamin E and Se (vitamin E 10 mg + Na₂SeO₃ 0·2 mg over a day). This administration was done for 5 weeks. Blood samples were taken from animals at the end of the dosage period and biochemical parameters in serum samples and haematological parameters in total blood were determined. The levels of total cholesterol (p<0·01) and number of white blood cells (p<0·001) in blood were significantly higher in the vitamin E group than in the control group. The levels of ALP, total cholesterol (p<0·01) and number of white blood cells (p<0·01) in blood were significantly higher in the selenium group than in the controls. The levels of glucose (p<0·05), ALP (p<0·01), total cholesterol (p<0·001) and number of white blood cells (p<0·01) were higher in the vitamin E and selenium combined group than in the controls. Other parameters considered within this trial (ALT, LDH, creatinine, albumin, total protein, amylase, creatine kinase, HDL, triglycerides, total lipid, sodium, chloride, uric acids, red blood cell, haemoglobin, packed cell volume, MCV, MCH, MCHC) did not show statistically significant differences between the control and injected groups. The results indicated that blood glucose and total cholesterol levels, ALP activity and white blood cell counts were significantly increased by intraperitoneal administration of vitamin E and selenium in rats. Copyright © 1999 John Wiley & Sons, Ltd.     

Protective role of intraperitoneally administered vitamin E and selenium in rats anesthetized with enflurane

The aim of this investigation was to determine levels of liver vitamins A and E and blood biochemical and hematological parameters in the enflurane anesthesia of rats. Fifty adult male Wistar rats were used in this study. All rats were randomly divided into five groups. The first and second groups were used as the control and anesthesia control groups, respectively, and only the placebo was intraperitoneally injected. The third group was intraperitoneally administered with vitamin E (dl/-α-tocopheryl acetate, 100 mg/kg body weight), the fourth group with Se (Na₂SeO₃ 1.5 mg/kg body weight), and the fifth group with vitamin E and Se (dl-α-tocopheryl acetate, 100 mg/kg body weight + Na₂SeO₃ 1.5 mg/kg body weight). This administration was done for three times with overday intervals and the second, third, forth, and fifth group rats were taken to enflurane anesthetise for 2 h. The liver vitamin E level was slightly lower in the anesthesia control group than in control group. However, the liver vitamin E content was significantly (p < 0.05 andp < 0.01) increased in vitamin E, Se, and combination groups, whereas the vitamin A level in liver was not statistically different. In general, plasma levels of alanine aminotransferase, creatin kinase, total bilirubin, urea, red blood cell counts, packet cell volume, and hemoglobulin values were significantly (p < 0.05 andp < 0.001) increased during the anesthesia and returned to near control values after the vitamin E plus selenium injection. However, administration of vitamin E had less effect on the hematological and biochemical parameters compared to that of selenium and their combination with vitamin E. However, the white blood cell count and levels of alkaline phosphatase, aspartate aminotransferase, total cholesterol, triglycerides, total protein, and creatinine were not statistically influenced by the anesthesia. In conclusion, we observed that plasma levels of some enzymes and metabolites were significantly increased in the enflurane anesthesia of rats, whereas the liver vitamin E levels were slightly decreased. Therefore, we observed that vitamin E and selenium have a protective effect against anesthesia complication, but the effect of selenium appears to be much greater than the vitamin E.     

Effect of vitamin A pretreatment on Escherichia coli-induced lipid peroxidation and level of 3-nitrotyrosine in kidney of guinea pig

In the present study, we report the effect of vitamin A (Vit A, retinol palpitate) on kidney lipid peroxidation and 3-nitrotyrosine (3-NT) levels induced after Escherichia coli administration to guinea pigs. Vit A was administrated intraperitoneally (i.p.) to guinea pigs at a dose 15,000 IU/kg per day for 7 days prior to E. coli injection. On day 8, the animals were injected i.p. with E. coli dosed at 12 ×10⁹ colony forming units per kilogram. Kidneys were collected 6 h after administration of E. coli. Malondialdehyde (MDA) as a lipid peroxidation product, and 3-NT levels were measured by reverse phase high-performance liquid chromatography. There was a significant increase in MDA and 3-NT levels in lipopolysaccaharide-induced group (p<0.001). 3-NT was not detectable in kidney of normal control animals. However, Vit A administration prior to E. coli injection prevented 3-NT formation but did not prevent the rice in MDA level of kidney (p<0.001). Vit A alone did not alter the MDA level in the kidney of the control group. (Mol Cell Biochem 278: 33–37, 2005)     

Vitamin E in immunity and reproductive performance in pigs

This review is focused on studies of vitamin E in immunity and reproductive performance in pigs. There are reports that vitamin E can have a positive effect on some parameters of the immune system in pigs. The optimal level of vitamin E needed to improve the immune system has not been determined because of several factors such as the composition of the diet, feed consumption, the rate of animal growth and living conditions or stress. Moreover, the way of action of vitamin E in enhancing immunity is still unclear but according to reports it may have antioxidant properties as well as an immunomodulator effect. In several studies, an increase in litter size and a reduction of preweaning piglet mortality have resulted from increasing dietary vitamin E intake during gestation or by intramuscular injection of vitamin E and/or selenium. However, according to reports, the positive effect of vitamin E on reproductive performance remains unclear due to the low number of animals used in most experiments.