Comparison of oxidative stress and the frequency of polymorphisms in the HFE gene between hemoglobin S trait blood donors and sickle cell disease patients

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Protective effect of nickel chloride on superoxide damage: enhancement of CuZn superoxide dismutase affinity to the oxygen free radical.

The effect of nickel from soluble NiCl2 on Cu-Zn superoxide dismutase (SOD) activity, as well as on rate of nitro blue tetrazolium reduction, was studied in vitro since lipid peroxidation has been implicated in cell damage by nickel insoluble compounds, whose toxicity and carcinogenicity are well established. The physical and chemical nature of nickel compounds is one of the key determinations of its toxicity. Soluble nickel freely enter cells, but is just as readily excreted reducing the opportunity for production of lipid damage. Nickel from NiCl2 strongly activated SOD activity. In vitro addition of nickel chloride to a crude lung preparation altered the KM for SOD without changing the Vmax. Nickel chloride produced increased enzyme affinity to the substrate, because decreased (O2-) concentration that yields half-maximal velocity. The combination of nickel and SOD may contribute to stabilization of the particular conformation of SOD responsible for maximal catalytically activity.

Increased oxygen radical and high-dietary-carbohydrate pancreatic damage.

These data suggest that an improved understanding of the relationship between high dietary carbohydrate and the rate of lipid peroxidation may give some insight into possible treatment modalities for pancreatic damages and may shed light on molecular mechanisms underlying certain pathological processes. High dietary carbohydrate lesions are age related and induced alterations on ceruloplasmin, phospholipids, total proteins, copper and zinc serum levels. Significantly increased serum and pancreatic amylase, and lipoperoxide determinations were observed in 20 month old rats. Cu-Zn superoxide dismutase was decreased in these animals. Daily injection of Cu-Zn superoxide dismutase conjugated with polyethylene glycol (SOD-PEG) prevented the serum and pancreatic changes, indicating that superoxide radical is an important intermediate to high dietary carbohydrate lesion.

Protective effect of safranine on the mitochondrial damage induced by Fe(II)citrate: Comparative study with trifluoperazine

In this study, we show that safranine at the concentrations usually employed as a probe of mitochondrial membrane potential significantly protects against the oxidative damage of mitochondria induced by Fe(II)citrate. The effect of safranine was illustrated by experiments showing that this dye strongly inhibits both production of thiobarbituric acid-reactive substances and membrane potential decrease when energized mitochondria were exposed to Fe(II)citrate in the presence of Ca 2+ ions. Similar results were obtained with the lipophylic compound trifluoperazine. It is proposed that, like trifluoperazine, safranine decreases the rate of lipid peroxidation due to its insertion in the membrane altering the physical state of the lipid phase.

Toxic mechanism of nickel exposure on cardiac tissue

The incidence of cardiovascular disease has increased in the general population, and cardiac damage is indicated as one important cause of mortality. In addition, pollution and metal exposure have increased in recent years. For this reason, toxic effects of metals, such as nickel, and their relation to cardiac damage should be urgently established. Although free radical-mediated cellular damage and reactive oxygen species have been theorized as contributing to the nickel mechanism of toxicity, recent investigations have established that free radicals may be important contributors to cardiac dysfunction. However, there is little information on the effect of nickel exposure on markers of oxidative stress in cardiac tissue. Nickel exposure (Ni2+ 100 mg L-1 from NiSO4) significantly increased lipoperoxide and total lipid concentrations in cardiac tissue. We also observed increased serum levels of cholesterol (59%), lactate dehydrogenase (LDH-64%), and alanine transaminase (ALT-30%) in study animals. The biochemical parameters recovered to the control values with tocopherol intake (0.2 mg 200 g-1). Vitamin E alone significantly decreased the lipoperoxide concentration and increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the heart. Since no alterations were observed in catalase and GSH-Px activities by nickel exposure while SOD activities were decreased, we conclude that superoxide radical (O2 -) generated by nickel exposure is of primary importance in the pathogenesis of cardiac damage. Tocopherol, by its antioxidant activity, decreased the toxic effects of nickel exposure on heart of rats.

Toxic mechanism of cadmium exposure on cardiac tissue

The presence of toxic substances in the workplace environment requires systematic evaluation of exposure and health status in exposed subjects. Cadmium is a highly toxic element found in water. Although free mediated cellular damage and reactive oxygen species (ROS), had been theorized as contributing to the cadmium mechanism of toxicity, and recent investigations have established that free radicals may be important contributors to cardiac dysfunction, there is little information on the effect of cadmium exposure on markers of oxidative stress in cardiac tissue. Cadmium exposure (Cd2+ - 100 mg/1-from CdCl2) in drinking water, during 15 days, significantly increased lipoperoxide and decreased the activities of superoxide dismutase and glutathione peroxidase. No alterations were observed in catalase activity in heart of rats with cadmium exposure. We also observed decreased glycogen and glucose concentration and increased total lipid content in cardiac tissue of rats with cadmium exposure. The decreased activities of alanine transaminase and aspartate transaminase reflected decreased metabolic protein degradation, and increased lactate dehydrogenase activity was related with increases in capacity of glycolysis. Since the metabolic pathways were altered by cadmium exposure, we can conclude that Cd2+ exposure induced ROS and initiate some series of events that occur in the heart and resulted in metabolic pathways alterations.

Avaliação do colesterol plasmático em coelhos com hipercolesterolemia induzida e tratados com extrato etanólico de própolis

The propolis (bee glue) is a product rich in flavonoids, which are known for antioxidant activities, a protective action to the lipoproteins LDL-cholesterol against lipid peroxidation. Because they have antioxidant properties, we investigated the effect of the ethanolic extract propolis on the plasma level of cholesterol in rabbits (Oryctolagus cuniculus) submitted to hypercholesterolaemia. The animals were divided into 4 groups. G 1=received commercial feed and water, G 2=received enriched feed and water, G 3=received enriched feed and ethanol, G 4=received enriched feed and ethanolic extract of propolis. The hypercholesterolaemia was induced with commercial feed enriched with egg yolk. The animals received the ethanolic extract propolis at the concentration of 100 mg/kg daily. Weekly, after fast of 14 hours, the samples of blood were collected from the marginal vein of the ear. The plasma was used for the estimation total cholesterol. From the results obtained, we verified that the ethanolic extract propolis significantly reduced the plasma level cholesterol (109,59 mg/dL, p<0,05), compared to the animals treated with ethanol (331,38 mg/dL), and also to those receiving the commercial feed only, with cholesterol at 269,74 mg/dL.

The effect of chilling on the photosynthetic activity in coffee (Coffea arabica L.) seedlings. The protective action of chloroplastid pigments

Coffea arabica is considered to be sensitive to low temperatures, being affected throughout its entire life cycle. Injury caused by chilling (low temperatures above zero degree centigrade) is characterized primarily by inhibition of the photosynthetic process. The objective of this work was to evaluate the role of photosynthetic pigments in the tolerance of coffee (C. arabica L.) seedlings to chilling. The evaluation the photosynthetic activity was made by emission of Chl a fluorescence at room temperature (25°C) in vivo and in situ, using a portable fluorometer. The pigment content was obtained by extraction with 80% acetone, while estimation of membrane lipid peroxidation was determined by measuring the MDA content in leaf tissue extracts. The results indicated a generalized reduction in the quantum yield of PSII when the seedlings were maintained in the dark. The reduction occurred in the seedlings submitted to chilling treatment as well as in the control ones. This demonstrates that not only chilling acts to cause an alteration in PSII. It is possible that the tissue storage reserves had been totally exhausted, with the respiratory rate exceeding the photosynthetic rate; the later was nil, since the seedlings were kept in the dark. The efficiency in the capture, transfer and utilization of light energy in PS11 photochemical reactions requires a sequence of photochemical, biochemical and biophysical events which depend on the structural integrity of the photosynthetic apparatus. However, this efficiency was found to be related to the protective action of chloroplastid pigments, rather than to the concentration of these pigments.

Comparative study of multiple dosage of quercetin against cisplatin-induced nephrotoxicity and oxidative stress in rat kidneys

Quercetin, a typical bioflavonoid ubiquitously present in fruits and vegetables, is considered to be helpful for human health. Cisplatin (cDDP) is one of the most active cytotoxic agents in the treatment of a wide range of solid tumors. The aim of this study was to investigate the possible effect of quercetin, a bioflavonoid with antioxidant potential, on cisplatin-induced nophrotoxicity and lipid peroxidation in rats. Gavage administrations of water, propylene glycol and quercetin (50 mg/kg) were made 24 and 1 h before saline or cDDP (5 mg/kg) ip injections and were repeated daily for 2, 5 or 20 subsequent days. Rats were killed 2, 5 and 20 days after ip injections, and blood and urine samples were collected to determine plasma creatinine, urine volume and osmolality. The kidneys were removed to determine the levels of thiobarbituric acid-reactive substances (TBARS) and for histological studies. Cisplatin increased lipid peroxidation, urine volume and plasma creatinine levels and decreased urine osmolality. Treatment with quercetin attenuated these alterations. These results demonstrate the role of oxidative stress and suggest a protective effect of quercetin on cisplatin-induced nephrotoxicity in adult Wistar rats. Copyright © 2006 by Institute of Pharmacology Polish Academy of Sciences.

Remodelação cardíaca induzida pelo tabagismo (tabaquismo)

Previous studies accomplished by our group suggest that tobacco smoke exposure results in cardiac remodeling, with progressive ventricular dysfunction. However, the mechanisms involved in this process are not known. Therefore, our laboratory has been trying to identify the potentials responsible mechanisms for cardiac remodeling induced by tobacco. The blood pressure increase, the renin-angiotensin system and the oxidative stress can modulate this process. On the other hand, the activation of MMP-2 or MMP-9, as well as lipid peroxidation, don't seem to participate of the morphologic and functional alterations induced by tobacco smoke exposure.

Efeito hipocolesterolêmico dos ácidos caféicos da própolis

Natural products have been used in tratamed of hypercholesterolemia. The bee products have been promoting up the man's interest, among them it stands out the propolis, coleted by bees, rich in polyphenols. The biggest polyphenols of propolis was flavonoids and caffeic acids, which have antioxidant power, presenting the protective action to the lipoprotein LDL-cholesterol against the lipid peroxidation. Therefore, the present work was evaluated whether the caffeic acids of the Botucatu's propolis (Botucatu-SP) affect the levels of plasma cholesterol, in rabbits submitted to the rich diet in cholesterol. The animals were divided in three groups: C (n=2) they received commercial ration and water for the whole period; S (n=2) they received normal ration and water in the first period and supplemented ration and water in the second and third periods; S+T (n=5) they received normal ration and water in the first period, supplemented ration and water in the second period and supplemented ration and extract rich in caffeic acids in the third period. The caffeic acids were administered diluted in water, being in the concentration of 0.05g caffeic acid mL -1 kg -1 of animal. Weekly, after fast of 14 hours, the samples of blood were collected from the marginal vein of the ear for determination of the plasma levels of total cholesterol and their fractions. The caffeic acids of the propolis reduced the plasma concentration of total cholesterol in 30% (280 for 199 mg dL -1) in the rabbits treated with flavonoids, while in the animals of the group S those levels were reduced discreetly (380 for 400 mg dL -1). The animals of the group C maintained this biochemical parameter in the normality range during the whole period (50 mg dL -1). Therefore, we concluded that the caffeic acids exert inhibititory activity in the metabolism of the cholesterol, being considered as a substance of action against the hypercholesterolemia.

Intestinal anti-inflammatory activity of coumarin and 4-hydroxycoumarin in the trinitrobenzenesulphonic acid model of rat colitis

Coumarins represent an important class of phenolic compounds with multiple biological activities, including inhibition of lipidic peroxidation and neutrophil-dependent anion superoxide generation, anti-inflammatory and immunosuppressor actions. All of these proprieties are essential for that a drug may be used in the treatment of inflammatory bowel disease. The present study examined intestinal anti-inflammatory activity of coumarin and its derivative, the 4-hydroxycoumarin on experimental ulcerative colitis in rats. This was performed in two different experimental settings, i.e. when the colonic mucosa is intact or when the mucosa is in process of recovery after an initial insult. The results obtained revealed that the coumarin and 4-hydroxycoumarin, at doses of 5 and 25 mg/kg, significantly attenuated the colonic damage induced by trinitrobenzenesulphonic acid (TNBS) in both situations, as evidenced macroscopically, microscopically and biochemically. This effect was related to an improvement in the colonic oxidative status, since coumarin and 4-hydroxycoumarin prevented the glutathione depletion that occurred as a consequence of the colonic inflammation. © 2008 Pharmaceutical Society of Japan.

Lipoperoxidação, perda de viabilidade celular e diminuição da liberação de IL-8 de neutrófilos humanos na presença de corpos cetônicos

Type-1 diabetes patients suffer from frequent episodes of acidosis caused by an increased fatty acid metabolism and consequently increased plasma level of acetoacetate (AcAc) and β-hydroxybutyrate (β-HOB). This article describes a study of the effects of pathological concentrations of AcAc and β-HOB on lipoperoxidation, cell viability and the release of the CXCL8 (IL-8) cytokine by activated neutrophils. Neutrophils from healthy donors were isolated by density gradient (Histopaque® 1077/1119) and incubated with the ketone bodies. Lipoperoxidation was determined as thiobarbituric acid reactive substances (TBARS). The cell viability was evaluated by the release of intracellular lactate dehydrogenase. The release of CXCL8 was measured by ELISA in a 24-h culture of opsonized zymosan-stimulated neutrophils. AcAc, but not β-HOB, provoked a dose-dependent increase in the neutrophil membrane lipoperoxidation (p<0.05; r =0.9915). In the cytotoxicity assay, a dose-dependent release of LDH was observed when the neutrophils were incubated with AcAc in concentrations up to 40 mM (p<0.05). β-HOB was devoid of effect. The release of CXCL8 was inhibited by AcAc and β-HOB in a dose-dependent manner. In conclusion, these results suggest that the accumulation of ketone bodies in diabetic patients could be involved in their usually increased susceptibility to infection.

Biochemical biomarkers in Nile tilapia (Oreochromis niloticus) after short-term exposure to diesel oil, pure biodiesel and biodiesel blends

Fossil fuels such as diesel are being gradually replaced by biodiesel, a renewable energy source, cheaper and less polluting. However, little is known about the toxic effects of this new energy source on aquatic organisms. Thus, we evaluated biochemical biomarkers related to oxidative stress in Nile tilapia (Oreochromis niloticus) after two and seven exposure days to diesel and pure biodiesel (B100) and blends B5 and B20 at concentrations of 0.01 and 0.1mLL -1. The hepatic ethoxyresorufin-O-deethylase activity was highly induced in all groups, except for those animals exposed to B100. There was an increase in lipid peroxidation in liver and gills in the group exposed to the higher concentration of B5. All treatments caused a significant increase in the levels of 1-hydroxypyrene excreted in the bile after 2 and 7d, except for those fish exposed to B100. The hepatic glutathione-S-transferase increased after 7d in animals exposed to the higher concentration of diesel and in the gill of fish exposed to the higher concentration of pure diesel and B5, but decreased for the two tested concentrations of B100. Superoxide dismutase, catalase and glutathione peroxidase also presented significant changes according to the treatments for all groups, including B100. Biodiesel B20 in the conditions tested had fewer adverse effects than diesel and B5 for the Nile tilapia, and can be suggested as a less harmful fuel in substitution to diesel. However, even B100 could activate biochemical responses in fish, at the experimental conditions tested, indicating that this fuel can also represent a risk to the aquatic biota. © 2011 Elsevier Ltd.