Maria A. Pavanato

Hepatoprotective effects of pecan nut shells on ethanol-induced liver damage

The hepatoprotective activity of the aqueous extract of the shells of pecan nut was investigated against ethanol-induced liver damage. This by-product of the food industry is popularly used to treat toxicological diseases. We evaluated the phytochemical properties of pecan shell aqueous extract (AE) and its in vitro and ex vivo antioxidant activity. The AE was found to have a high content of total polyphenols (192.4±1.9 mg GAE/g), condensed tannins (58.4±2.2 mg CE/g), and antioxidant capacity, and it inhibited Fe(2+)-induced lipid peroxidation (LP) in vitro. Rats chronically treated with ethanol (Et) had increased plasmatic transaminases (ALT, AST) and gamma glutamyl transpeptidase (GGT) levels (96%, 59.13% and 465.9%, respectively), which were effectively prevented (87; 41 and 383%) by the extract (1:40, w/v). In liver, ethanol consumption increased the LP (121%) and decreased such antioxidant defenses as glutathione (GSH) (33%) and superoxide dismutase (SOD) (47%) levels, causing genotoxicity in erythrocytes. Treatment with pecan shell AE prevented the development of LP (43%), GSH and SOD depletion (33% and 109%, respectively) and ethanol-induced erythrocyte genotoxicity. Catalase activity in the liver was unchanged by ethanol but was increased by the extract (47% and 73% in AE and AE+Et, respectively). Therefore, pecan shells may be an economic agent to treat liver diseases related to ethanol consumption.

Resveratrol improves sperm motility, prevents lipid peroxidation and enhances antioxidant defences in the testes of hyperthyroid rats.

Hyperthyroidism may lead to a loss of sperm motility and an increase in oxidative stress (OS) in testes and may cause male reproductive disorders. Thus, the use of compounds with antioxidant properties may be a strategy for preventing these disorders. The effect of resveratrol (RSV) on sperm motility and on variables of the antioxidant status in the testes of rats with triiodothyronine-induced hyperthyroidism (100μg/kg) was investigated. Hyperthyroid rats presented lower sperm motility, higher levels of lipid hydroperoxides and thiobarbituric reactive substances, lower catalase and glutathione peroxidase activities and higher glutathione-S-transferase activity in their testes than control animals. RSV treatment (1mg/kg and 10mg/kg) was able to prevent these effects in the hyperthyroid rats and had no effect in the control animals. In conclusion, RSV might be a strategy for therapeutic intervention to preserve sperm motility and to prevent OS in testes, preserving testicular function in those with hyperthyroidism.

Bioaccumulation and oxidative stress parameters in silver catfish (Rhamdia quelen) exposed to different thorium concentrations

The objective of this study was to evaluate the effect of chronic thorium (Th) exposure on bioaccumulation, metabolism (through biochemical parameters of the muscle) and oxidative parameters (lipidic peroxidation levels and antioxidant enzymes in the gills and in the hepatic and muscular tissues) of silver catfish (Rhamdia quelen). Silver catfish juveniles were exposed to different waterborne Th levels (in microg L(-1)): 0 (control), 25.3+/-3.2, 80.6+/-12.0, 242.4+/-35.6, and 747.2+/-59.1 for 30 d. The gills and skin were the organs that accumulated the highest Th levels. The increase in the waterborne Th concentration corresponded to a progressive increase in the Th levels in the gills and kidney. Chronic Th exposure causes alterations in the oxidative parameters of silver catfish gills, which are correlated with the Th accumulation in this organ. The levels of GST decreased in the gills of fish exposed to 747.2 microg L(-1) Th and SOD activity decreased in silver catfish exposed to 242.4 and 747.2 microg L(-1) Th. In addition, the increase in the LPO in the gills exposed to 242.4 and 747.2 microg L(-1) Th suggests that higher oxidative damage occurred in the gills. However, in the liver and muscle, these alterations occurred mainly in the lowest waterborne Th level. Metabolic intermediates in the muscle were altered by Th exposure, but no clear relationship was found.

Biochemistry, cytogenetics and bioaccumulation in silver catfish (Rhamdia quelen) exposed to different thorium concentrations.

The objective of this study was to evaluate the effect of thorium (Th) bioaccumulation on the metabolism of silver catfish (Rhamdia quelen) through biochemical parameters of the muscle (glycogen, glucose, lactate, protein, and ammonia). In addition, lipidic peroxidation levels (TBARS), catalase (CAT) and glutathione-S-transferase (GST) in the gills and in hepatic and muscular tissues were also analyzed. Cytogenetic parameters were studied through the evaluation of nuclear abnormalities in red blood cells. Silver catfish juveniles were exposed to different waterborne Th levels (in microg L(-1)): 0 (control), 25.3+/-3.2, 69.2+/-2.73, 209.5+/-17.6, and 608.7+/-61.1 for 15 days. The organs that accumulated the highest Th levels were the gills and skin. The increase of waterborne Th concentration corresponded to a progressive increase of Th levels in the gills, liver, skin and kidneys, with the highest accumulation in the gills and skin. Metabolic intermediates in the muscle were altered by Th exposure, but no clear relationship was found. CAT and GST activities in the hepatic and muscular tissues of this species suggest that the enzymatic activities can be stimulated at the lowest Th levels and inhibited at the higher levels (mainly in 608.7 microg L(-1)). The results of the cytogenetic assay contribute to this hypothesis because the higher toxicity in blood samples was found in juveniles exposed to 69.2 and 209.5 microg L(-1) Th.

Effects of parboiled rice diet on oxidative stress parameters in kidney of rats with streptozotocin-induced diabetes.

The effect of parboiled rice (PR) and white rice (WR) diets on oxidative stress (OS) parameters was investigated in the kidneys of rats with streptozotocin-induced diabetes (40 mg kg(-1), iv). The experimental groups (n=8) were control fed with PR (CPR), control fed with WR, diabetic fed with PR, and diabetic fed with WR. After 30 days of treatment, all animals were anesthetized and exsanguinated before removal of kidneys, which were used to determine thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides, carbonyl protein, superoxide dismutase, catalase, glutathione peroxidase (GPx), glutathione reductase, glutathione-S-transferase activities, and levels of glutathione (GSH). Total phenolic compounds were determined in WR and PR grains. Our data indicated that diabetes induced increase in TBARS and lipid hydroperoxides levels. Although PR has not prevented the rise in the levels of these measurements, its consumption by our animals resulted in higher GPx activity and GSH content than that of the CPR. Moreover, PR also presented concentration of total phenolic compounds 127% higher than WR grains. Thus, its consumption in this diabetic condition is suggested because this seems to confer greater protection against OS in the renal tissue of diabetic animals.

Oxidative stress parameters in silver catfish (Rhamdia quelen) juveniles infected with Ichthyophthirius multifiliis and maintained at different levels of water pH

The aim of this study was to determine oxidative stress parameters in the liver, gill and muscle of silver catfish juveniles infected with Ichthyophthirius multifiliis and maintained at pH 5.0 or 7.0 for three days. Juveniles were infected by adding one I. multifiliis-infected juvenile and water containing theronts to tanks. After the appearance of white spots on the skin, infected juveniles exposed to pH 5.0 and 7.0 showed significantly higher thiobarbituric acid reactive substances (TBARS) levels in the liver and gills compared to uninfected juveniles. Liver of infected juveniles exposed to pH 7.0 showed higher catalase (CAT) and lower glutathione-S-transferase (GST) activities, but those maintained at pH 5.0 showed significantly higher GST activity than uninfected juveniles. The gills of infected juveniles showed significantly higher CAT (day two) and GST activity at both pH 5.0 and 7.0 compared to uninfected juveniles. Muscle of infected juveniles showed significantly lower CAT and GST activity and TBARS levels (at day three) when maintained at both pH 5.0 and 7.0 compared to uninfected juveniles. In conclusion, I. multifiliis infection induces liver and gill damage via lipid peroxidation products in silver catfish, but higher antioxidant enzyme activity could indicate a greater degree of protection against this parasite.

Effect of N-acetylcysteine on the spinal-cord glutathione system and nitric-oxide metabolites in rats with neuropathic pain.

Since N-acetylcysteine (NAC) is a donor of cysteine, we studied the relationship between NAC and concentration of oxidized and reduced glutathione (GSH/GSSG ratio), and glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activities in the lumbosacral spinal cord of rats with chronic constriction injury (CCI) of the sciatic nerve that received NAC (150mg/kg/day, i.p.) or 0.9% saline solution for 3 or 10 days. Hydrogen peroxide (H2O2) and nitric-oxide (NO) metabolites were also measured. Von Frey hair and hot-plate tests showed hyperalgesia at day 1 in CCI rats. Hyperalgesia persisted at all other times in saline-treated CCI rats, but returned to pre-injury values in NAC-treated CCI rats after 3 postoperative days. GST activity and the GSH/GSSG ratio increased in saline-treated CCI rats, while the NAC treatment increased GST and GPx activities at day 10, with no significant change in the GSH/GSSG ratio. NAC treatment did not affect H2O2 levels, but it reduced NO metabolites in CCI rats 3 days after the surgery. Thus, the anti-hyperalgesic effect of NAC appears not to involve its action as a cysteine precursor for GSH synthesis, but involves a decrease in NO.

Myrcia sylvatica essential oil mitigates molecular, biochemical and physiological alterations in Rhamdia quelen under different stress events associated to transport

The effects of pre-transport handling and addition of essential oil of Myrcia sylvatica (EOMS) during transport on stress pathways activation in Rhamdia quelen were investigated. Fish (n=400, 25.2±2.9g) were captured in production ponds and transferred to 100-L tank (density 100g L-1). After 24h, 10 fish were sampled (before transport group). The remaining fish were placed in plastic bags (n=30 or 32 fish per bag, density 150g L-1) containing 5L of water (control), ethanol (315μLL-1, vehicle) or EOMS (25 or 35μLL-1), in triplicate, transported for 6h and sampled (n=10 animals per group). Indicators of stress and metabolism, as well as mRNA expression of brain hormones were evaluated. Previously, full-length cDNAs, encoding specific corticotropin-releasing hormone (crh) and proopiomelanocortins (pomca and pomcb), were cloned from whole brain of R. quelen. Crh expression increased after 24h of capture and handling, whereas cortisol and glucose plasmatics enhanced their values in the control group. Transport with EOMS reduced plasma cortisol and lactate levels, while ethanol and EOMS groups increased Na+/K+-ATPase gill activity compared to control. Gene expression of crh, pomcb, prolactin and somatolactin mRNAs were lower after transport with EOMS compared to control. EOMS was able to mitigate the stress pathways activation caused by transport, maintaining a balance in body homeostasis. Thus, EOMS is recommended as sedative in procedures as transport and the pre-transport handling requires greater attention and use of tranquilizers.

Chronic aspartame intake causes changes in the trans-sulphuration pathway, glutathione depletion and liver damage in mice

No-caloric sweeteners, such as aspartame, are widely used in various food and beverages to prevent the increasing rates of obesity and diabetes mellitus, acting as tools in helping control caloric intake. Aspartame is metabolized to phenylalanine, aspartic acid, and methanol. Our aim was to study the effect of chronic administration of aspartame on glutathione redox status and on the trans-sulphuration pathway in mouse liver. Mice were divided into three groups: control; treated daily with aspartame for 90 days; and treated with aspartame plus N-acetylcysteine (NAC). Chronic administration of aspartame increased plasma alanine aminotransferase (ALT) and aspartate aminotransferase activities and caused liver injury as well as marked decreased hepatic levels of reduced glutathione (GSH), oxidized glutathione (GSSG), γ-glutamylcysteine (γ-GC), and most metabolites of the trans-sulphuration pathway, such as cysteine, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH). Aspartame also triggered a decrease in mRNA and protein levels of the catalytic subunit of glutamate cysteine ligase (GCLc) and cystathionine γ-lyase, and in protein levels of methionine adenosyltransferase 1A and 2A. N-acetylcysteine prevented the aspartame-induced liver injury and the increase in plasma ALT activity as well as the decrease in GSH, γ-GC, cysteine, SAM and SAH levels and GCLc protein levels. In conclusion, chronic administration of aspartame caused marked hepatic GSH depletion, which should be ascribed to GCLc down-regulation and decreased cysteine levels. Aspartame triggered blockade of the trans-sulphuration pathway at two steps, cystathionine γ-lyase and methionine adenosyltransferases. NAC restored glutathione levels as well as the impairment of the trans-sulphuration pathway.

Oxidative stress parameters in juvenile Brazilian flounder Paralichthys orbignyanus (Valenciennes, 1839) (Pleuronectiformes: Paralichthyidae) exposed to cold and heat shocks

The aim of this study was to determine oxidative stress parameters in the liver and gill of Brazilian flounder juveniles (307.0 ± 16.0 g and 30.0 ± 4.0 cm) submitted to different water temperature (17.1, 23.0 and 28.8oC) for 72 h and maintained at salinity 25‰. After the acclimation of 7 days, in 23oC, fish were transferred to 200 L tanks containing seawater (salinity 25‰) at 28.8oC (heat shock), 17.1oC (cold shock) or 23.0oC (control), five replicates (five fish tank-1). The sampled collection occurred in 0 (pre-challenge), 3, 24, 48 and 72 h after temperature shock. Flounder exposed to 17.1oC and 28.8oC showed significantly higher TBARS levels and GST activity in the liver post-exposition (PE) in relation to the control (23oC). CAT activity in liver present a significantly increase at 17.1oC, in first 48 h, and subsequently decrease in 72 h PE in relation to 28.8oC. The gills of flounder showed significantly higher TBARS levels, GST and CAT activity when submitted at 17.1 and 28.8oC in relation to 23.0oC. There were observed changes in lipid peroxidation levels (LPO), CAT and GST activities in the liver and gill of Brazilian flounder in response to reactive oxygen species (ROS) produced by thermal shocks.