Chronic iron overload induces gender-dependent changes in iron homeostasis, lipid peroxidation and clinical course of experimental autoimmune encephalomyelitis.

To analyze iron- and gender-dependent mechanisms possibly involved in pathogenesis of multiple sclerosis (MS) in this study we evaluated the effects of iron overload (IO) on iron status and lipid peroxidation processes (LPO) in tissues of female and male DA rats during chronic relapsing experimental autoimmune encephalomyelitis, a well-established MS animal model. Rats were treated by iron sucrose (75mg/kg bw/day) or with saline solution during two weeks before the sensitization with bovine brain homogenate in complete Freund's adjuvant. Clinical signs of EAE were monitored during 29 days. Serum and tissues of CNS and liver were sampled before immunization and at day 13th post immunization (during acute phase of EAE). The determination of ferritin, iron, malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) and evaluation of histopathology were performed by ELISA, ICP spectrometry and immunohistochemistry. Results showed that IO in female EAE rats accelerated the onset of disease. In contrast, in male rats it accelerated the progression of disease and increased the mortality rate. During acute phase of EAE female IO rats sequestered more Fe in the liver, spinal cord and in the brain and produced more ferritin than male EAE rats. Male rats, however, reacted on IO by higher production of MDA or 4-HNE in the neural tissues and showed greater signs of plaque formation and gliosis in spinal cord. The data point to sexual dimorphism in mechanisms that regulate peripheral and brain iron homeostasis and imply that men and women during MS might be differentially vulnerable to exogenous iron overload.

Preexposure to Olive Oil Polyphenols Extract Increases Oxidative Load and Improves Liver Mass Restoration after Hepatectomy in Mice via Stress-Sensitive Genes.

Polyphenols can act as oxidants in some conditions, inducing redox-sensitive genes. We investigated the effect of preexposure to the olive oil polyphenols extract (PFE) on time-dependent changes in the hepatic oxidative state in a model of liver regeneration-a process in which oxidative stress associated with the metabolic overload accounts for the early events that contribute to the onset of liver self-repair. Liver regeneration was induced by one-third hepatectomy in mice. Prior to hepatectomy, mice were intraperitoneally given either PFE (50 mg/kg body weight) or saline for seven consecutive days, while respective controls received vehicle alone. Redox state-regulating enzymes and thiol proteins along with the mRNA levels of Nrf2 gene and its targets γ-glutamylcysteine synthetase and heme oxygenase-1 were determined at different time intervals after hepatectomy. The liver mass restoration was calculated to assess hepatic regeneration. The resulting data demonstrate the effectiveness of preexposure to PFE in stimulating liver regeneration in a model of a small tissue loss which may be ascribed to the transient increase in oxidant load during the first hours after hepatectomy and associated induction of stress response gene-profiles under the control of Nrf2.

Influence of cadmium on metallothionein expression and products of lipid peroxidation in the organs of hares (Lepus europaeus Pallas).

Cadmium occurs naturally in the environment and as an anthropogenic pollutant. Exposure to low concentrations of cadmium is inevitable and may produce toxic effects. Another important aspect of cadmium toxicity is its interaction, often antagonistic, with essential elements such as selenium. The aim of this study was to highlight the risks of long-term exposure to low cadmium concentrations, using a scientific and chemical approach and hares (Lepus europaeus Pallas) as model organisms in a field study. Two study areas were monitored. Levels of cadmium and selenium were quantified in the organs of hares, the expression of metallothioneins I + II and the products of lipid peroxidation were determined. The median cadmium concentrations (wet weight) in the muscle, liver, kidney and brain of hares from an exposed group ranged from 0.033 to 0.037, 0.763 to 1.054, 3.090 to 16.594 and 0.016 to 0.087 µg g(-1), respectively; whereas, the median selenium concentrations (wet weight) ranged from 0.100 to 0.108, 0.153 to 0.332, 0.677 to 0.701 and 0.078 to 0.116 µg g(-1), respectively. Expression of the metallothioneins I + II proteins was observed in tissues. Lipid peroxidation (LPO) levels, measured as malondialdehyde (MDA) equivalents, increased with the cadmium concentration. Further research on long-term exposure to low concentrations of cadmium in the environment is needed.

Metallothioneins I/II expression in rat strains with genetically different susceptibility to experimental autoimmune encephalomyelitis.

OBJECTIVES: Compared to the Dark Agouti (DA), the Albino Oxford (AO) rat strain exhibits lower susceptibility to the induction of experimental autoimmune encephalomyelitis (EAE). Here, we investigated the potential contribution of the heavy metal-binding proteins metallothioneins (MTs) I/II to these effects. METHODS: Rats were immunized with bovine brain homogenate emulsified in complete Freund's adjuvant or only with complete Freund's adjuvant. The expression patterns of MTs mRNA and proteins and tissue concentrations of Zn2+ and Cu2+ were estimated in the brain and in the liver on days 7 and 12 after immunization, by real-time PCR, immunohistochemistry and inductively coupled plasma spectrometry, respectively. Additionally, the hepatic transforming growth factor beta and nuclear factor kappa B immunoreactivities were tested. RESULTS: Clinical signs of EAE were not induced in AO rats, but they upregulated the expression of MT I/II proteins in the brain (hippocampus and cerebellum) and in the liver, similarly as DA rats. The transcriptional activation of MT-I occurred, however, only in DA rats, which accumulated also more zinc in the brain and in the liver. In contrast, intact AO rats had greater hepatic MT-I mRNA immunoreactivity and more Cu2+ in the hippocampus. Besides, in immunized AO rats a high upregulation of transforming growth factor beta and nuclear factor kappa B immunoreactivities was found in several hepatic structures (vascular endothelium, Kupffer cells and hepatocytes). CONCLUSIONS: Our data show that AO and DA rats differ in constitutive and inductive MT-I gene expression in the brain and in the liver, as well as in the hepatic cytokine profile, suggesting that these mechanisms may contribute to the discrepancy in the susceptibility to EAE.

Effect of olive oil on brain's lipid and calcium content after partial hepatectomy in mice.

The aim of this study was to investigate the influence of olive oil (00) enriched diet on the lipid content of mice brain during the early phase of liver regeneration and to test a relationship of these changes with calcium content. C57BI mice were fed over 21 days with diet enriched with olive oil, containing predominantly oleic acid (18:1n-9). The animals were one-third partially hepatectomised (pHx) under aether anaesthesia. Total lipids were extracted from tissue samples with a chloroform-methanol (2:1, v/v) mixture according to Folch et al. Mineral concentration was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES) after microwave brain tissue digestion. The diet containing 00 increased both total lipid content and the calcium concentration in brain during the early phase of liver regeneration (12hrs post pHx), suggesting that monounsaturated oleic acid might interact with some metal-dependent activities that control changes in the brain during liver regeneration.

Kinetics of tissue iron in experimental autoimmune encephalomyelitis in rats.

To elucidate the role of iron in the pathomechanisms of autoimmune CNS disorders, we estimated the tissue concentrations of Fe(2+) in the brain, spinal cord, and liver in the chronic relapsing form of experimental autoimmune encephalomyelitis (EAE). The disease was induced in Dark Agouti (DA) strain of rats, by subcutaneous injection of bovine brain homogenate in complete Freund's adjuvant (CFA). Control rats consisted of unsensitized rats and of rats treated with CFA or saline. The data obtained by clinical assessment and by inductively coupled plasma spectrometry have shown that the attacks of disease (on the 12th and 22nd post-immunization day) were followed by high accumulation of iron in the liver. Additionally, during the second attack of disease, the decreased concentration of Fe(2+) was found in cervical spinal cord. The data point to regulatory effects of iron and hepatic trace elements regulating mechanisms in the pathogenesis of EAE.

Differential effect of high dietary iron on alpha-tocopherol and retinol levels in the liver and serum of mice fed olive oil- and corn oil-enriched diets.

The influence of dietary fats on cellular alpha-tocopherol and retinol uptake in iron overload is poorly understood. The aim of this study was to investigate the effect of a high-iron diet on the retinol and alpha-tocopherol levels in mice fed olive oil- and corn oil-enriched diets. Mice were fed for 3 weeks a standard mouse chow (the control group) and diets enriched with 5% by weight of corn oil or olive oil. Diets of the mice fed corn oil and olive oil were additionally supplemented with 1% by weight carbonyl iron. Both dietary oils and iron increased the liver iron uptake. High-iron feeding induced oxidative stress in mice liver, measured as a thiobarbituric acid-reactive substance level. Both fats and iron induced changes in the liver fatty acid composition. Liver retinol and alpha-tocopherol stores increased with iron supplementation in the olive oil-enriched diet, with a simultaneous decrease in serum. The results suggest that the influx of alpha-tocopherol and retinol from serum to the liver is induced by high dietary iron. This redistribution appears to be stronger for retinol than for alpha-tocopherol and is also higher in mice fed olive oil than in mice fed corn oil, suggesting that the composition of dietary lipids is important in the treatment of high-iron tissue conditions. In conclusion, the results of this study suggest that the increase of hepatic alpha-tocopherol and retinol levels in the olive oil-based diet is a dietary-dependent responsive mechanism that probably is not primarily related to an increased risk of oxidative damage induced by high-iron intake.

Oxidative stress in mice: effects of dietary corn oil and iron.

The aim of this study was to analyze the effect of high dietary Fe on liver antioxidant status in mice fed a corn-oil-enriched diet. Male Balb/c mice were fed for 3 wk with a standard diet enriched with 5% by weight of corn oil with adequate Fe (FCO diet) or supplemented with 1% carbonyl Fe (FCOFe diet). The control group was fed a standard diet. The high-Fe diet induced a twofold increase of hepatic Fe level. However, an increase of thymic Fe level has been induced solely by dietary fat. The hepatic copper (Cu) level slightly decreased in the FCO diet. In the spleen, the high-Fe diet-induced increase of Fe level was negatively correlated with the Cu level. The antioxidant status was influenced by both dietary fat and Fe. Mice fed corn-oil-enriched diets had a higher concentration of thiobarbituric acid-reactive substances (TBARS), with a greater increase in the FCOFe diet. Fatty acid analysis showed decreased n-3 and n-6/n-3 ratio, particularly in the FCOFe diet. Hepatic Cu/Zn superoxide dismutase (CuZn-SOD) activity was decreased in FCO diet, and Fe supplementation caused a further decrease in the enzyme activity. These results suggest that feeding with corn oil-enriched diet increases oxidative damage by decreasing antioxidant enzyme defense. The high-Fe diet additionally affects the antioxidant defense system, further increasing the tissue's susceptibility to lipid peroxidation. Additionally, both corn-oil- and Fe-enriched diets have increased the Cu requirement in mice.

Liver fatty acid and element changes after partial hepatectomy in mice fed olive oil- and corn oil-enriched diets.

The purpose of this study was to explore the effect of dietary fats on the hepatic fatty acid profile of mice liver after partial hepatectomy and to observe a correlation with changes in element content. Male Balb/C mice were divided into three groups: the control animals fed a standard diet (FSD), the FOO group fed a diet enriched with 5% olive oil, and the FCO group fed a diet enriched with 5% corn oil. Hepatic fatty acid and element content were analyzed within each group in intact animals and on d 1, 2, and 7 after partial hepatectomy. During the regenerative process, proportions of 18:1n-9 and 18:3n-6 substantially increased in the FSD diet, correlating with the Zn level. On the other hand, 20:4n-6 and 22:6n-3 decreased on d 1 and 2. Consequently, a significant increase in the n-6: n-3 ratio was found in these animals. In the FOO diet, a decreased polyunsaturated fatty acid/monounsaturated fatty acid (PUFA/MUFA) ratio was related to a significant decrease in PUFA content, mainly the result of decreased 20:3n-6 and 20:4n-6. The proportion of 18:1n-9 was highly increased when compared with other diets and remained high during the regeneration. Furthermore, the n-6: n-3 ratio was increased on d 2 and 7. Zn increased on d 1 and 2, and Fe increased on d 2. Feeding with corn oil generally induced an increase in the PUFA n-6 series, compared with other diets. The PUFA n-3 series decreased and the 18:1n-9 increased on d 1, compared to intact animals. Consequently, the n-6: n-3 ratio was elevated during the regeneration. Zn increased on d 1 and 2, whereas Fe remained unchanged until d 7, when it decreased. Decreased 20:4n-6 on d 1 and 2, as well as Cu on d 7, and increased Zn in the first 2 d were common to all three diets. These findings suggest that some significant signals transmitted during the regenerative process have induced alterations in the fatty acid composition and changes in the liver element content, which can be modified by the diet.