Antioxidant effects of metronidazole in colonic tissue.

Reactive oxygen species, primarily generated in the mitochondria, contribute to tissue injury in inflammatory bowel diseases. The efficacy of metronidazole (MTZ) in some situations of inflammatory bowel disease may result not only from its antibiotic effect, but also from an antioxidant effect. We evaluated, under physiologic conditions, the antioxidant potential of MTZ on the biomarkers of oxidative damage to proteins (protein carbonyls), lipids (malondialdehyde), and the levels of antioxidant defense (glutathione) in the small bowel, large bowel, and liver of control and MTZ-treated rats. Basal levels of protein carbonyls and malondialdehyde were significantly higher in the colon compared with those in the small intestine, whereas glutathione levels were quite similar along the bowel. MTZ reduced significantly the colonic oxidative damage to proteins without any side effect in the liver. This is the first evidence of an antioxidant effect of MTZ on oxidative protein damage in the colon under physiologic conditions.

Zinc and intestinal anaphylaxis to cow's milk proteins in malnourished guinea pigs.

Zinc supplementation could favor recovery from diarrhea in malnourished children. As the recent experimental evidence suggests that oxidative stress and intestinal anaphylaxis may contribute to the intestinal dysfunction associated with malnutrition, we postulated that zinc could act through antioxidant or antianaphylactic properties. Control (C), malnourished (M), and malnourished zinc-treated (MZ) guinea pigs were, respectively, fed a normal 30% protein diet, a low 4% protein diet, and a low 4% protein diet plus 1800 ppm of zinc. Milk proteins were included in the diets to trigger intestinal anaphylaxis. Milk sensitization was assessed by passive cutaneous anaphylaxis (PCA) against beta-lactoglobulin and by intestinal anaphylaxis measured in Ussing chambers by the increase in short circuit-current after addition of beta-lactoglobulin (deltaIsc(betaLg)). Oxidative stress was assessed by intestinal lipid peroxidation. The intestinal secretion was assessed by deltaIsc induced by inflammatory mediators. Malnutrition increased the level of anti-betaLg reaginic antibodies [PCA = 1.19 +/- 0.79 and 0.69 +/- 0.67 log(l/titer) in M versus C guinea pigs, p = 0.07] and enhanced intestinal anaphylaxis (deltaIsc(betaLg)) = 16.4 +/- 9.9 and 9.1 +/- 5.8 microA/cm2 in M versus C guinea pigs, p = 0.07), without inducing intestinal lipid peroxidation. Moreover, malnutrition enhanced significantly the intestinal secretory response to histamine and 5-hydroxytryptamine. Administration of pharmacologic doses of zinc during malnutrition inhibited the increase in milk sensitization induced by malnutrition, both at the systemic [PCA = 0.35 +/- 0.55 log(l/titer) in MZ guinea pigs, p = 0.03 versus M] and intestinal (deltaIsc(betaLg)) = 2.8 +/- 2.5 microA/cm2 in MZ guinea pigs; p = 0.001 versus M) level, and prevented the hypersecretion in response to histamine and 5-hydroxytryptamine. These data suggest that zinc has antianaphylactic and antisecretory properties that may contribute to its capacity to prevent intestinal dysfunction during malnutrition.

Endogenous glutathione as potential protectant against free radicals in the skin of vitamin A deficient mice.

In order to evaluate the relationships between oxidative degradation of lipids and antioxidant defence systems in the skin, weanling female SKH1 hairless mice were randomly divided into two groups. Each group was fed a well-balanced diet, supplemented, in one group with 5 IU vitamin A/g, and vitamin A free in the other, for 20 wk. Liver and plasma vitamin E were increased in mice fed the vitamin A-free diet. Superoxide dismutases, catalase and Se-glutathione peroxidase were determined in dorsal skin homogenates, as well as the concentration of reduced glutathione (GSH) and of thiobarbituric acid-reactive substances (TBARS); the latter is an index of peroxidation of murine skin cell membranes. Vitamin A deficiency did not alter enzyme activities but enhanced the skin reserve of GSH, which appeared to be the reason for a decrease in endogenous lipid peroxidation. Statistical analysis showed a highly significant negative correlation (R2 > 0.6) between the concentrations of TBARS and GSH in these untreated animals. GSH could play a critical role in maintaining a lower background of lipid alteration in the skin of healthy animals and minimizing individual risk.

Modulation of ultraviolet light-induced oxidative stress in mice skin related to dietary vitamin A and selenium intake.

Weanling female SKH1 hairless mice were randomly divided into 4 groups. Each group was fed a particular regimen for 20 weeks: 1) normal basal diet with 5 IU vitamin A/g and 0.45 microgram selenium/g, 2) vitamin A deficient, 3) selenium deficient, and 4) vitamin A plus selenium deficient. Three hours before being sacrificed, half of the animals were subjected to UV A + B irradiation (3 J/cm2). Superoxide dismutases (SOD), catalase, Se glutathione peroxidase activities were determined in dorsal skin homogenates, as well as the concentrations of GSH and of thiobarbituric acid-reactive substances (TBARS), the latter being an index of lipid peroxidation. Ultraviolet light altered the antioxidant defense of mouse skin tissue: GSH level, catalase and Se glutathione peroxidase activities were lowered and SOD was unequally enhanced according to the nutritional status. Vitamin A and Se deficiencies did not perceptibly aggravate the UV-induced oxidative stress, although the former enhanced the decline of catalase expression induced by the irradiation. Probably through an adaptive mechanism, both dietary deficiencies increased the skin reserve of antioxidant GSH and thus appear to modulate the effects caused by reactive oxygen species.

Rat serum osteocalcin concentration is decreased by restriction of energy intake.

We studied the effects of energy restriction on serum osteocalcin concentration and bone formation rate in rats. The experiment was designed to achieve energy restriction by reducing the carbohydrate intake while providing identical quantities of protein, fat, vitamins and minerals. Energy intakes of three groups of post-weaning male rats were restricted by 20, 40 and 60% for 4 wk. Serum calcium, phosphorus, transthyretin, triiodothyronine (T3), thyroxine (T4), 1,25-dihydroxycholecalciferol, 25-hydroxycholecalciferol and immunoreactive parathyroid hormone (iPTH) concentrations were determined. Energy restriction (20, 40 and 60%) produced a significant and gradual drop of serum osteocalcin concentrations, although the serum concentrations of its key regulators, i.e., 1,25-dihydroxycholecalciferol and iPTH, were not significantly affected. On the contrary, serum concentrations of calcium, phosphorus, transthyretin, T3 and T4 were significantly lower in the energy-restricted groups. However, our results do not support their implication in the regulation of serum osteocalcin synthesis by energy intake. Serum osteocalcin concentration was positively correlated with bone mineral apposition (r = 0.50, P < 0.05) and bone mineralization (r = 0.50, P < 0.05) rates suggesting that its decrease resulted from a reduction of bone formation, and not from abnormal mineralization, because osteoid seam thickness was not modified. Energy intake seems to be an important determinant of serum osteocalcin concentration and bone formation; however, the exact mechanism underlying this regulation remains to be determined.

Excessive dietary selenium decreases the vitamin A storage and the enzymatic antioxidant defence in the liver of rats.

This study investigated the influence of selenium intake, over 8 weeks, on vitamin A level and on enzymatic antioxidant defence in the liver of young rats. Deficient animals were fed a well-balanced diet but without selenite addition; the Se content of this diet which originated from natural Se content of ingredients was 0.05 mg/kg. Controls were fed the same diet with 0.40 mg/kg added Se. The two other groups received high levels of Se, 2.05 or 4.05 mg/kg. Excessive Se intake decreased the concentrations of retinol and retinyl palmitate in the liver. The linear regression analysis indicated a significant (P < 0.001) dose-dependent vitamin A decline. As expected, Se deficit lowered glutathione peroxidase activity. The highest Se excess decreased the enzymatic antioxidation: Zn,Cu superoxide dismutase, catalase, glutathione peroxidase activities. Data showed that high dietary Se can sometimes enhance carcinogenesis and our results suggest that it is best to be cautious in administrating Se to humans with the aim of preventing diseases.

Oxidative stress may contribute to the intestinal dysfunction of weanling rats fed a low protein diet.

Intestinal function is impaired in malnutrition. Because oxidative stress is a component of gastrointestinal injury, and malnutrition may reduce antioxidant defenses, we investigated the involvement of oxidative stress in the intestinal dysfunction due to malnutrition. Weanling rats were fed either a normal protein (22% casein) or a low protein (6% casein) diet for 4 wk. In intestinal homogenates, we assessed free radical damage and enzymatic antioxidant defenses. In jejunal fragments mounted in Ussing chambers, we measured ionic transport by short-circuit current (Isc) and protein permeability by transepithelial fluxes of beta-lactoglobulin. Catalase activity and the thiobarbituric acid-reacting substances concentration were greater in intestinal mucosa of the low protein group, whereas the glutathione concentration and the activities of superoxide dismutase and Se-dependent glutathione peroxidase were the same as in the normal protein group intestinal mucosa. Both basal Isc and the delta Isc induced by glucose and forskolin, as well as beta-lactoglobulin fluxes, were higher in the low protein group. Exogenous H2O2 stress increased Isc significantly more in the low protein than the normal protein group but did not alter protein permeability. These results show that malnutrition induces both intestinal free radical damage and altered epithelial transport, suggesting that oxidative stress may contribute to the intestinal dysfunction associated with malnutrition.

Effects of protein deficiency on lipid peroxidation in the small intestine and liver of rats.

This study investigated the influence of protein restriction on lipid peroxide content (thiobarbituric acid-reactive substances) and the intracellular antioxidant defence system in the small intestine and the liver. Weanling male Sprague-Dawley rats were divided into two groups: a low-protein group fed ad lib. a diet containing 6% casein for 4 wk, and a control group fed a diet containing 22% casein but restricted to the same dietary intake as that of the low-protein group. Compared with pair-fed controls, the protein-deficient rats exhibited a decrease in glutathione content in the small intestine and the liver. Thiobarbituric acid-reactive substances were increased in the intestine only. Data on the enzymatic antioxidant defence system in the protein-deficient animals showed an increase in catalase activity in the intestine but a decrease of this activity in the liver. The activity of selenium-dependent glutathione peroxidase was decreased in the liver and remained unchanged in the intestine. Superoxide dismutase was not modified by protein deficiency in either tissue. In control rats, enzyme activities were 6-43 times higher in the liver than in the intestine. The deleterious effects of protein deficiency appeared more marked in the intestine.

Dietary restriction decreases thiobarbituric acid-reactive substances generation in the small intestine and in the liver of young rats.

This study investigated the influence of dietary restriction on thiobarbituric acid-reactive substances (TBARS) contents and on the intracellular antioxidant defence system in the small intestine or liver of young rats. Four weeks of diet restriction (-40%) lowered the TBARS level in both organs. No variations were found for the superoxide dismutase and catalase activities; only liver seleno-dependent glutathione peroxidase was enhanced by the restriction. The protection appeared more marked in the intestine than in the liver, and would be dependent on glutathione concentration.

Effect of prototypic polychlorinated biphenyls on hepatic and renal vitamin contents and on drug-metabolizing enzymes in rats fed diets containing low or high levels of retinyl palmitate.

Two groups of weanling male Sprague-Dawley rats fed a diet supplemented with either 0.6 or 6 retinol equivalents/g diet were each separated into three further groups receiving 300 mumol 2,2',4,4',5,5'-hexachlorobiphenyl/kg body weight, 300 mumol 3,3',4,4'-tetrachlorobiphenyl kg/body weight or vehicle only (corn oil). Only the coplanar (3,4)2Cl congener caused a slight reduction in food intake, thymic atrophy and led to a significant decrease in the liver vitamin A storage. The vitamin A lost by the liver was approximately the same in both dietary groups; however an increased renal accumulation of vitamin A was observed in the high vitamin A group. Serum retinol was reduced by (3,4)2Cl treatment but remained unchanged by (2,4,5)2Cl exposure. Total amounts of ascorbic acid and its oxidation products were increased in the liver and in the kidney by both xenobiotics while niacin and thiamine concentrations were lowered by (3,4)2Cl only. Microsomes from vitamin A-deficient rats exhibited a marked decrease in the anisotropy parameter. After (2,4,5)2Cl exposure, an increase in membrane fluidity was observed linked to a decrease in cholesterol/phospholipid (C/P) ratio. Treatment with (3,4)2Cl caused a significant decrease in the index of fluorescence polarization only in the low vitamin A group even if the C/P ratio was enhanced in both dietary groups. This study shows that the polychlorinated biphenyl with the 3-methylcholanthrene-type pattern of induction of cytochrome P-450 has more profound effects on B group vitamins and particularly vitamin A homeostasis than does the phenobarbital-type inducer. Moreover, this situation, which has been found to be similar to that in vitamin A deficiency, is not ameliorated by a high dietary vitamin A intake.

Effect of phenoclor DP6 on enzyme-altered foci and lipid peroxidation in livers of aflatoxin B1-initiated rats.

This study investigates the capacity of phenoclor DP6 to promote aflatoxin B1 (AFB1)-induced putative preneoplastic foci in the liver. In male Sprague-Dawley rats pretreated with AFB1 (ip injection of 1 or 2 mg/kg body weight once weekly for 3 consecutive wk) and given a diet containing 50 ppm phenoclor DP6 for 11 days, the number of area of putative liver preneoplastic lesions were increased approximately four-fold as indicated by the number and gamma-glutamyl transpeptidase activity of enzyme-altered foci; this change was also accompanied by an increase in hepatic microsomal lipid peroxidation and a decrease in Se-glutathione peroxidase and superoxide dismutase activities. The results indicate that phenoclor DP6 exerts a promoting effect in AFB1-initiated rats.

Lipid peroxidation of rat liver microsomes membranes related to a protein deficiency and/or a PCB treatment.

In this study, we investigated the influence of protein deficiency on lipid peroxidation (LP) and cellular defense systems against oxidative damage in control or polychlorinated biphenyl (PCB) treated rats. Rats were fed either a standard diet (22% casein) or a low protein diet (3.5% casein) for 1, 2 and 6 weeks. Five days prior to killing, one half of the animals were given a single i.p. injection of Phenoclor DP6 (50 mg/kg body weight). In protein deficient rats, liver vitamin E was depressed and ascorbate level was lowered. Total and selenium-dependent glutathione peroxidases (GSH-Px) activities were decreased whereas glutathione reductase (GSH-red) was enhanced. Enzymatic and non enzymatic LP ('spontaneous' or with ADP-Fe2+) were increased. Phenoclor DP6 treatment enhanced liver ascorbate concentration. Microsomal LP was increased. Total and selenium-GSHPx remained unmodified while GSH-red was increased. Liver glutathione and alpha-tocopherol contents appeared to be independent of the PCB injection. Our data suggest that low protein intake and PCB exposure may reduce liver defensive protection against electrophilic species.

The effects of vitamin A nutritional status on microsomal lipid peroxidation and alpha-tocopherol level in rat liver.

In vitamin A-deficient rats, liver glutathione peroxidase activity was decreased, alpha-tocopherol content was strongly enhanced, but microsomal liquid peroxidation remained unchanged.

The effect of dietary imbalances on the activation of benzo[a]pyrene by the metabolizing enzymes from rat liver.

Male Sprague-Dawley rats (70-80 g) were fed ad libitum a standard control diet (22% casein, 5% lard), or a high lipid diet (30% lard) or a low protein diet (6% casein) or a standard diet containing 50 ppm phenoclor DP6. After 6 weeks on these diets, the cytochrome P-450 microsomal content, the benzo[a]pyrene monooxygenase (BaP-MO) and the epoxide hydrolase (EH) were assayed. The formation of mutagenic B(a)P metabolites which covalently bind with DNA was compared. The activity of BaP-MO and of EH were increased by the high lipid diet (+27% and 106% respectively) and by the phenoclor DP6 treatment (+63% and 400% respectively), compared to the standard diet. In animals fed a low protein diet the BaP-MO was decreased (-34%) and the EH activity was strongly increased (+262%) compared to those fed a standard diet. All experimental diets increased both the activation of BaP to metabolites able to bind DNA and the mutagenicity of BaP versus TA98 Salmonella typhimurium strain. It was concluded that dietary imbalances can be considered as a factor in chemical carcinogenesis.

[Protein restriction and metabolism of xenobiotics]. / Restriction protéique et métabolisme des xénobiotiques.

Young male rats were fed a balanced diet or a low-protein diet (casein 6%; w/w) for 6 weeks. After 2 weeks, each group was divided, one half being maintained as the control and the other half being fed diet-containing (Phenoclor DP6) at a level of 50 mg/kg for 4 weeks. A low-protein diet lowered the liver glutathione content and the activity of the mono-oxygenases. It lowered the inductive effect of PCB towards P450, NADPH-cytochrome c reductase, aminopyrine demethylase and aniline hydroxylase, but it raised the benzo(a) pyrene hydroxylase induction.

[Polychlorobiphenyl (Phenoclor DP6) and the metabolism of xenobiotics: effect of a high lipid diet]. / Polychlorobiphenyle (phenoclor DP6) et metabolisme des xenobiotiques: effets d'un regime hyperlipidique.

Young male rats were fed a balanced diet or a hyperlipidic diet (lard 30%, w/w) for 6 wk. After the first 2 wk each group was divided, one half being maintained as the control and the other half being fed the same diet with a polychlorinated biphenyl (Phénoclor DP6) added at a level of 50 mg/kg diet for 4 wk. In animals fed the balanced diet, DP6 increased the microsomal-protein and cytochrome P-450 content and the specific activity of the monooxygenases. The high-fat diet increased the inductive effect of DP6 on cytochrome P-450, aminopyrine demethylase and benzo[a]pyrene hydroxylase, but not on aniline hydroxylase or cytochrome c reductase.

Lipid composition and monooxygenase relationship in liver microsomal membranes from rats fed unbalanced diets.

The cholesterol and phospholipid content of microsomal membrane from rats fed either a high lipid (H1) (30% lard) or a low protein (Lp) diet (6% casein) have been compared with those from rats fed a standard (St) diet (22% casein, 5% lard). For each diet, half of the group was treated with Phenochlor DP6. A significant increase in the phospholipid cholesterol ratio was observed in rats fed a high lipid diet or treated with DP6. These effects tend to increase the microsomal membrane fluidity. The protein deficiency decreased the phospholipid/cholesterol ratio and then the fluidity of the endoplasmic reticulum membrane. The specific activity of cytochrome P-450 to hydroxylate aniline which is independent to the enzymic form of cytochrome P-450 was closely correlated with the viscosity status of the microsomal membrane.