Effect of dietary selenium on the induction of altered hepatic foci and hepatic tumors by the peroxisome proliferator ciprofibrate

The purpose of this study was to determine if the dietary antioxidant selenium could inhibit hepatocarcinogenesis induced by peroxisome proliferators, which are hypothesized to induce tumors by increased production of hydrogen peroxide or other reactive oxygen species. Rats were fed diets containing the peroxisome proliferator ciprofibrate and one of three concentrations (0.04, 0.2, or 1.0 ppm) of selenium for 6 or 21 months. The incidence of hepatic tumors and the number and volume of gamma-glutamyl transpeptidase-positive, ATPase-negative, glucose-6-phosphatase-negative, and glucose-6-phosphatase-positive foci at 21 months were lower in rats fed higher levels of selenium (no foci or tumors were seen at 6 mo). Indices of oxidative damage in the liver (thiobarbituric acid reactants, conjugated dienes, and lipid-soluble fluorescence products), however, were not decreased in rats fed the high-selenium diet. Therefore, selenium was protective against ciprofibrate-induced hepatocarcinogenesis, but not by reducing the degree of oxidative damage. The liver selenium and glutathione concentrations, and liver selenium-dependent glutathione peroxidase activity, increased as dietary selenium increased. Therefore, inhibition of carcinogenesis by selenium was correlated with increased levels of glutathione and glutathione peroxidase, but these did not inhibit the indices of oxidative damage. Peroxisomal beta-oxidation also increased with the dietary selenium content; it therefore does not appear to be a factor in the inhibition of hepatocarcinogenesis in rats fed higher levels of selenium.     

Dietary selenium intake controls rat plasma selenoprotein P concentration

The purpose of this study was to determine the effect of dietary selenium on selenoprotein P concentration. Selenoprotein P was quantitated in plasma by radioimmunoassay. Selenium-dependent glutathione peroxidase activity in plasma and liver 105,000 x g supernatant was measured for comparison. Weanling male rats were fed a selenium-deficient diet or a control diet that contained 0.5 mg selenium/kg as Na2SeO4. The concentration of selenoprotein P fell at approximately the same rate in the rats fed the selenium-deficient diet as did plasma glutathione peroxidase activity. Groups of weanling rats were fed different levels of selenium for 8 wk. Selenoprotein P concentration was proportional to dietary selenium level up to 0.1 mg/kg and was a greater percentage of control values than was glutathione peroxidase activity. No increment in selenoprotein P concentration occurred between 0.1 and 0.5 mg selenium/kg diet. These results indicate that the concentration of selenoprotein P in the plasma is directly dependent on selenium supply in the diet up to 0.1 mg/kg. There is overlap between the dietary selenium ranges in which selenoprotein P concentration and glutathione peroxidase activity increase, but the selenoprotein P range is lower than the glutathione peroxidase range.     

Platelet glutathione peroxidase activity as an index of selenium status in rats

Glutathione peroxidase activity in platelets increased stepwise in selenium-depleted rats that were repleted with graded levels of dietary sodium selenite. In a 3-phase depletion/repletion/depletion feeding study, glutathione peroxidase activity was similar in platelets and liver, which apparently contains the largest labile pool of selenium in the body. The activity of glutathione S-transferase (selenium-independent glutathione peroxidase) in platelets was low and was not affected by selenium deficiency, even though hepatic transferase was markedly elevated in selenium-deficient rats. Vitamin E deficiency did not affect activities of glutathione peroxidase or glutathione S-transferase in platelets or liver. Determination of glutathione peroxidase activity in platelets apparently is a promising technique for assessing selenium status and, possibly, for measuring selenium bioavailability.     

Effect of selenium depletion and repletion on plasma glutathione and glutathione-dependent enzymes in the rat

Selenium deficiency has several known biochemical effects. In the rat, these effects include loss of glutathione peroxidase (GSH-Px) activity, increased plasma glutathione concentration and increased liver glutathione S-transferase (GSH S-Tr) activity. The time course of the development of these changes in rats fed selenium-deficient diets and the time course of reversal of these changes in selenium-deficient rats fed graded levels of selenium were determined. As selenium deficiency was produced, liver cytosolic and plasma GSH-Px activities decreased first and were less than 5% of control when plasma glutathione concentration and liver GSH S-Tr activity began to increase. Elevated liver GSH S-Tr activity in selenium-deficient rats was corrected by refeeding selenium at the lowest level of supplementation (0.015 ppm) for 4 wk. GSH-Px activity required a supplementation of 0.10 ppm selenium for correction to control levels in 4 wk. Based on these studies a classification of the severity of selenium deficiency into mild, moderate and severe categories is proposed. In addition, the effect of dietary sulfur amino acid supplementation on plasma glutathione concentration was studied.     

High dietary intake of sodium selenite induces oxidative DNA damage in rat liver

One mechanism for the cancer-chemopreventive effects of high selenium (Se) intake is hypothesized to be antioxidant protection of DNA. In this work we examine DNA oxidation in whole animals as a function of dietary Se intake and carcinogen administration. Weanling male Sprague-Dawley rats were fed a basal, Torula yeast-based, Se-deficient diet supplemented with 0, 0.15, or 2.0 ppm Se as sodium selenite for 10 wk. They were then injected with 0, 0.1, or 10 mg /kg body weight of the pro-oxidant carcinogen N-nitrosodiethylamine. High levels of carcinogen and high levels of selenite intake each increased concentration of 8-hydroxy-2'-deoxyguanosine in liver DNA. Se-dependent glutathione peroxidase I gene expression and enzyme activity were dramatically reduced by dietary Se deficiency but were unaffected by carcinogen administration. There were no significant main or interactive effects of Se or carcinogen on activity or gene expression of the DNA repair enzyme 8-oxoguanine glycosylase I. These results do not support the hypothesis that high Se intake may be cancer-preventive by inhibiting oxidative DNA damage. Rather than inhibiting oxidative DNA damage, these findings suggest that high dietary intake of inorganic Se may promote in vivo DNA oxidation.     

Effect of dietary selenium and vitamin E on the antioxiant defense systems of rat erythrocytes.

The effect of dietary selenium and vitamin E on the important cellular antioxidant defense systems was studied in rat erythrocytes. Weanling male Sprague-Dawley rats were fed a basal selenium and vitamin E deficient diet and supplemented with either none or 0.5 ppm selenium and either none or 45 ppm vitamin E for 35 or 40 days. Depletion of dietary selenium resulted in marked decrease of glutathione (GSH) peroxidase in the red cells, but the levels of GSH, catalase and superoxide dismutase were not significantly altered. The red cells of rats fed the basal diet deficient in both selenium and vitamin E had significantly lower levels of GSH and GSH peroxidase, but not of catalase and superoxide dismutase, than in those fed the basal diet and supplemented with either selenium, vitamin E or both. The results suggest that depletion of dietary selenium and vitamin may have a precipitate effect on lowering the levels of GSH and GSH peroxidase in rat erytyrocytes.     

Effect of dietary restriction on the age-dependent changes in the expression of antioxidant enzymes in rat liver

The effects of aging and dietary restriction on the expression of several enzymes (superoxide dismutase, catalase and glutathione peroxidase) that are involved in free radical detoxification were studied in liver tissue from male Fischer F344 rats. The expression (i.e., activities and mRNA levels) of superoxide dismutase (Cu-Zn) and catalase decreased with age in liver. Dietary restriction (40% restriction of energy intake) increased the activities of superoxide dismutase (24 to 38%) and catalase (64 to 75%) in liver at 21 and 28 mo of age. Glutathione peroxidase activity in liver of diet-restricted rats was significantly higher (37%) at 28 mo of age than that of rats fed ad libitum. The age-related changes in the relative levels of mRNA for superoxide dismutase, catalase and glutathione peroxidase paralleled the changes in the activities of these enzymes in rats fed ad libitum or rats fed the restricted diet. Thus, the changes in the activities of superoxide dismutase, catalase and glutathione peroxidase with age and dietary restriction appear to arise from changes in the levels of mRNAs coding for these enzymes. Free radical damage, as measured by thiobarbituric acid-reactive material and lipofuscin accumulation, was lower in diet-restricted rats than in rats fed ad libitum.     

Glutathione peroxidase acitvity in intestinal and liver tissues of rats fed various levels of selenium, sulfur and alpha-tocopherol.

The effects of various levels of selenium, alpha-tocopherol and sulfur on glutathione peroxidase (GSH-Px) activity in intestinal and liver tissues were determined in male rats fed corn-soybean or Torula yeast diets. Rats fed corn-soybean diets had greater GSH-Px activity in the small intestine, colon and liver tissues, catalase activity and selenium in the liver, and body weight gains than those fed Torula yeast diets. GSH-Px activity in the small intestine, colon, and liver tissues as well as concentration of selenium in the liver increased with increasing levels of selenium in Torula yeast diets but not with corn-soybean diets. Tocopherol supplementation had no significant effect on GSH-Px activity in rats fed Torula yeast or corn-soybean diets supplemented with selenium. Supplemental sulfur decreased GSH-Px activity in the small intestine tissues and increased activity in colon tissues.     

Long-term consumption of a methionine-supplemented diet increases iron and lipid peroxide levels in rat liver

Methionine is a protective factor against various types of liver damage, but excessive dietary methionine is hepatotoxic. Because the mechanisms of L-methionine-related hepatotoxicity are poorly understood, the effect of long-term excessive L-methionine intake on the metabolism of iron and antioxidants was studied in rat liver to determine whether oxidative stress is involved. Wistar male rats were fed either an L-methionine-supplemented (16.0 g/kg) diet or a control diet for 1, 3, 6 and 9 mo. The growth rate of L-methionine-supplemented rats was significantly slower than that of controls. Iron, ferritin and thiobarbituric acid-reactive substances (TBARS) levels in the liver were greater in supplemented rats than in controls. Serum iron and transferrin levels were significantly lower in L-methionine-treated rats compared with controls. Serum ferritin did not differ between the two groups. Hepatic glutathione peroxidase activity, catalase activity and total glutathione concentrations were higher in rats fed the L-methionine-supplemented diet at 1 and 3 mo, but not at 6 and 9 mo. These results indicate that long-term consumption of excess L-methionine by rats may affect primarily iron metabolism rather than the antioxidant defense system and, consequently, induce an accumulation of iron.     

膳食蛋白质水平对硒利用的影响 Ⅰ、膳食蛋白质水平对大鼠血及组织中硒含量及谷胱甘肽过氧化物酶活力的影响

用低硒(0.03ppm)及高硒(0.30ppm)两类饲料喂养大鼠共三月,每类饲料蛋白质水平从5.3—13.4％,饲料基本成分为玉米和黄豆,实验结果发现,在不同硒水平下,饲料蛋白质水平对机体利用硒的影响大不相同。在低硒(0.03ppm)饲料下,低蛋白组动物血及血、心、肾及胰织组中的含硒量最高,饲养蛋白质水平愈高,各组织中硒含量则显著降低。及肝组织中的谷胱甘肽过氧化物酶活力有相同趋势。在高硒(0.30ppm)饲料组则无此种趋势,相反,蛋白质水平愈高,在肝脏中的贮存愈好。文中并讨论了在我国克山病区单纯补充硒的意义。     

Effects of dietary selenium and fish oil (MaxEPA) on arachidonic acid metabolism and hemostatic function in rats

This study investigated whether hemostatic function can be modified by both the consumption of fish oil and the level of dietary selenium. Male Sprague-Dawley rats were fed for 8 wk semipurified diets containing 7% corn oil (by wt) or 5.5% fish oil (MaxEPA) plus 1.5% corn oil with or without selenium supplementation. Consumption of the four diets caused no difference in weight gain, food intake or plasma malondialdehyde content. The selenium-supplemented rats had significantly higher levels of selenium and glutathione peroxidase activity in plasma. Fish oil feeding decreased ADP-induced platelet aggregation and increased bleeding time. The level of dietary selenium and type of oil interacted to influence the production of 6-keto-prostaglandin F1 alpha: more was produced when corn oil was fed in the selenium-deficient diets. These data suggest that the effect of dietary selenium on hemostatic function and the production of eicosanoids is minor.     

Oxidative stress and antioxidant status in rat blood, liver and muscle: effect of dietary lipid, carnitine and exercise

The purpose of this study was to determine the effect of dietary fat, carnitine supplementation, and exercise on oxidative damage and antioxidant status. Male Wistar rats (60 days old) were fed diets containing either hydrogenated fat (HF) or peanut oil (PO) with or without 0.5 mg % (of dry diet) carnitine. The rats were given exercise, i.e. swimming for 60 minutes, for 6 days/week for 6 months under each dietary condition. The blood malondialdehyde (MDA) level was higher in PO-fed rats, more so in exercising ones, while the same was not altered in carnitine-supplemented rats irrespective of the dietary fat or physical activity. The MDA level was significantly decreased in muscle, while increased in liver, of carnitine-fed rats. The blood glutathione (GSH) level also significantly increased in exercising rats as compared to sedentary ones, while carnitine supplementation elevated it in all the groups. Exercise and carnitine supplementation significantly lowered GSH levels in liver while increasing it in muscle. The glutathione peroxidase (GPX) activity was significantly increased in blood and muscle from PO-fed exercising rats as compared to sedentary ones, while carnitine supplementation elevated GPX activity in all the groups. The liver and muscle catalase (CAT) activities were significantly increased in PO-fed exercising rats, while carnitine did not have any effect. The pro-oxidative effect of the monounsaturated fatty acid (MUFA)-rich PO diet and prolonged regular exercise was less pronounced due to augmented antioxidant enzymes, GPX and CAT, induced by training to protect against the oxidative stress, while carnitine supplementation could help to counter lipid peroxidation due to exercise through redistribution of GSH from liver to blood and muscle.     

膳食维生素B_6对硒的生物利用率影响的研究 Ⅱ．对大鼠组织谷胱甘肽过氧化物酶活性的影响

实验用４周龄断乳雄性大鼠观察膳食维生素Ｂ６（ＶＢ６）对饲亚硒酸钠（ＳｅＬ）或ＤＬ－硒蛋氨酸（ＳｅＭｅｔ）大鼠组织中ＧＳＨ－Ｐｘ活性的影响，实验期为４周。实验证明：与补ＶＢ６各组相比，缺ＶＢ６各组血浆ＧＳＨ－Ｐｘ活性较高，而在红细胞中的结果相反（Ｐ＜０．０５）；缺ＶＢ６各组动物的骨骼肌、心肌和脾脏中ＧＳＨ－Ｐｘ活性都显著低于补Ｂ６各组（Ｐ＜０．０５），上述结果与给硒的化学形式无关。在用ＳｅＭｅｔ的处理组，缺Ｂ６大鼠的肝脏中ＧＳＨ－Ｐｘ活性显著低于补ＶＢ６的大鼠；而在用ＳｅＬ的处理组则没有观察到ＶＢ６的这种影响。本研究结果提示，硒掺入ＧＳＨ－Ｐｘ是通过一个ＶＢ６依存的过程。     

Dietary virgin olive oil protects against lipid peroxidation and improves antioxidant status in the liver of rats chronically exposed to ethanol

Excessive ethanol intake induces severe tissue damage particularly in the liver through the generation of reactive oxygen species. The aim of this study was to determine the effect of a virgin olive oil–rich diet on oxidative stress induced by chronic ethanol exposure in rats. Wistar rats were treated daily with a 35% ethanol solution for 6 weeks and fed with a standard chow or a diet containing 5% virgin olive oil. By administering ethanol to rats, a severe toxicity occurred in their liver, as assessed by the significantly elevated levels of serum transaminases. The hepatic malondialdehyde level, indicator of lipid peroxidation, was also increased in ethanol-treated rats, whereas the hepatic antioxidant enzyme activities, namely, superoxide dismutase, glutathione peroxidase, and catalase were significantly reduced. The activity of glutathione reductase remained unchanged in rats. Fatty acid composition of the liver was also significantly changed with ethanol intake. In contrast, virgin olive oil intake during ethanol treatment in rats resulted in a higher antioxidant activity and inhibited toxicity to the liver, as monitored by the reduction of transaminases levels and hepatic lipid peroxidation. Rats showed a better profile of the antioxidant system with normal glutathione peroxidase activity and ameliorated superoxide dismutase and catalase activities. In conclusion, results of this study indicate that olive oil ingestion by rats protects the liver from ethanol-induced oxidative damage by affecting the cellular redox potential.     

Influence of dietary methionine on the metabolism of selenomethionine in rats

To determine the influence of methionine on selenomethionine (SeMet) metabolism, weanling male rats were fed for 8 wk a basal diet marginally deficient in sulfur amino acids, containing 2.0 micrograms selenium (Se)/g as DL-SeMet and supplemented with 0, 0.3, 0.6 or 1.2% DL-methionine. Increased dietary methionine caused decreased selenium deposition in all tissues examined but increased glutathione peroxidase (GSHPx, EC 1.11.1.9) activity in testes, liver and lungs. A positive correlation was found between dietary methionine and the calculated percentage of selenium associated with GSHPx. In a second experiment, 75SeMet was injected into weanling male rats which had been fed the basal diet containing 2.0 micrograms selenium as DL-SeMet with or without the addition of 1.0% methionine. The selenoamino acid content of tissues and the distribution of 75Se in erythrocyte proteins were determined. In comparison to the rats fed the basal diet without added methionine, significantly more 75Se-selenocysteine was found in liver and muscle, more 75Se was found in erythrocyte GSHPx and less 75Se was found in erythrocyte hemoglobin of rats fed 1.0% methionine. These data suggest that methionine diverts SeMet from incorporation into general proteins and enhances its conversion to selenocysteine for specific selenium-requiring proteins, such as GSHPx.     

Effects of various dietary levels of selenium as selenite or selenomethionine on tissue selenium levels and glutathione peroxidase activity in rats

Weanling rats were fed a basal diet or this diet plus 0.2, 1.0, 2.0 or 4.0 mg/kg selenium (Se) as either selenite or selenomethionine (SeM). Except at the 0.2 mg/kg Se level, Se accumulated in all tissues at higher levels when SeM was fed than when selenite was given, and the magnitude of difference became more pronounced with increasing levels of dietary Se. This was particularly true for muscle and brain. Se levels in whole blood, testes, kidney and lungs were not significantly different between rats fed 0.2 mg/kg Se as selenite or as SeM, but the Se levels in liver, muscle and brain were higher in rats fed SeM. Although the tissue Se concentrations differed markedly, there were no differences in the glutathione peroxidase (GPX) activity in tissues of rats fed SeM rather than selenite. The percentage of Se associated with GPX was lower in all tissues from rats fed SeM than in those from rats fed selenite. These results indicate that the chemical forms of dietary Se can have a marked influence on biological responses, including bioavailability of dietary Se.     

Vitamin B6 dependence of selenomethionine and selenite utilization for glutathione peroxidase in the rat.

The biological availability of selenium from sodium selenite and selenomethionine for glutathione peroxidase activity was studied. Rats were fed ad libitum for 2 weeks a basal diet deficient in both selenium and vitamin B6, and then for the subsequent 2 weeks the same diet supplemented with vitamin B6 (2.5 micrograms as pyridoxine-HCl/g diet) or selenium (2 microgram/g diet) or both. In the presence of vitamin B6, selenite and selenomethionine increased equally the glutathione peroxidase activity in both the liver and erythrocytes above that of selenium-unsupplemented controls. In the absence of vitamin B6, selenomethionine was less effective in the liver and ineffective in the erythrocytes while selenite was equally effective in both tissues and was as effective as in the presence of vitamin B6. These results indicate that selenite selenium is readily available for glutathione peroxidase induction as compared with selenomethionine, and establish that vitamin B6 is involved in the metabolism of selenomethionine to supply selenium for glutathione peroxidase.     

Effects of dietary selenium on DMBA-induced carcinogenesis in rats fed a diet high in mixed fats

The effects of selenium intake on 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary carcinogenesis were examined in rats fed a diet high in mixed fats and representative of that consumed in North America. Six groups of 20 rats were fed an AIN-76 diet modified to contain 20% fat from lard:corn oil (3:1 wt/wt) and various amounts of selenium (0.1, 0.035, 0.1, 1.0, 2.0 or 4.0 mg Se/kg diet). At wk 5, animals in groups 2-6 were dosed with 4.32 mg of DMBA. Serum clinical parameters and the activities of plasma selenium-dependent and total glutathione peroxidase (GSHPx), erythrocyte GSHPx and superoxide dismutase (SOD) were determined every 4 wk for 25 wk. The extent of lipid peroxidation was determined by measuring urinary malondialdehyde during wk 13 and 24, and erythrocyte malondialdehyde at wk 25. Erythrocyte GSHPx was found to be a better indicator of selenium status than plasma activity, while SOD did not vary with dietary selenium. The group of animals fed 4.0 mg Se/kg diet had reduced numbers of tumors (P less than 0.01), but this reduction was associated with evidence of chronic selenium toxicity. Variations in GSHPx activity with dietary selenium did not result in differences in tumor incidence, nor in changes in lipid peroxidation in the other groups. Thus, nontoxic levels of selenium do not appear to offer any protective effect during carcinogenesis in rats fed a casein-based diet similar in fat content to that consumed by North Americans.     

Effects of α‐tocopherol,phenobarbital, and butylated hydroxyanisole during promotion of diethylnitrosamine‐initiated rat hepatocarcinogenesis

The promotion‐suppressing ability of two antioxidants was measured to determine the role of oxidative stress in hepatocarcinogenesis. Four‐day‐old female F344/N rats were dosed with diethylnitrosamine (10 mg/kg). After weaning, they were fed semipurified diets with and without 500 ppm α‐tocopherol, or the same two diets containing 500 ppm phenobarbital, or 5,000 ppm butylated hydroxyanisole (BHA) for 3 or 11 months. By 11 months, phenobarbital‐fed groups had eaten 30% more than other groups did (p < 0.05), suggesting a role for increased caloric intake in phenobarbital promotion. Phenobarbital and BHA significantly reduced body weights and increased liver weights compared with control rats. After three months, a‐tocopherol significantly suppressed mean volume of placental glutathione S‐transferase (PGST)‐positive altered hepatic foci (AHF), regardless of xenobiotic treatment. Phenobarbital increased and BHA decreased the numbers of AHF compared with those of the control group. After 11 months, mean focal volume was significantly suppressed by BHA compared with that of the control group, and phenobarbital increased the total volume of AHF [PGST‐positive plus y‐glutamyltransferase (GGT)‐positive AHF] compared with rats fed either control or BHA diets. BHA treatment also increased hepatic glutathione levels by 40% compared with control and rats fed phenobarbital.

In conclusion, a‐tocopherol had only a slight, early effect to suppress promotion of hepatocarcinogenesis. BHA suppressed some indices of promotion at both times and increased hepatic glutathione; however, BHA's toxicity (which suppressed body weight) may also be a factor in its supposable promotion‐inhibitory effects.