Greicy M. M. Conterato

Blood thioredoxin reductase activity, oxidative stress and hematological parameters in painters and battery workers: relationship with lead and cadmium levels in blood

Oxidative stress has been shown to be involved in lead and cadmium toxicity. We recently showed that the activity of the antioxidant enzyme thioredoxin reductase (TrxR) is increased in the kidneys of lead‐exposed rats. The present study evaluated the blood cadmium and blood lead levels (BLLs) and their relationship with hematological and oxidative stress parameters, including blood TrxR activity in 50 painters, 23 battery workers and 36 control subjects. Erythrocyte δ‐aminolevulinate dehydratase (δ‐ALA‐D) activity and its reactivation index were measured as biomarkers of lead effects. BLLs increased in painters, but were even higher in the battery workers group. In turn, blood cadmium levels increased only in the painters group, whose levels were higher than the recommended limit. δ‐ALA‐D activity was inhibited only in battery workers, whereas the δ‐ALA‐D reactivation index increased in both exposed groups; both parameters were correlated to BLLs (r = −0.59 and 0.84, P < 0.05), whereas the reactivation index was also correlated to blood cadmium levels (r = 0.27, P < 0.05). The changes in oxidative stress and hematological parameters were distinctively associated with either BLLs or blood cadmium levels, except glutathione‐S‐transferase activity, which was correlated with both lead (r = 0.34) and cadmium (r = 0.47; P < 0.05). However, TrxR activity did not correlate with any of the metals evaluated. In conclusion, blood TrxR activity does not seem to be a good parameter to evaluate oxidative stress in lead‐ and cadmium‐exposed populations. However, lead‐associated changes in biochemical and hematological parameters at low BLLs underlie the necessity of re‐evaluating the recommended health‐based limits in occupational exposure to this metal. Copyright

Prevention of unpredictable chronic stress-related phenomena in zebrafish exposed to bromazepam, fluoxetine and nortriptyline

RationaleSeveral model organisms have been employed to study the impacts of stress on biological systems. Different models of unpredictable chronic stress (UCS) have been established in rodents; however, these protocols are expensive, long-lasting, and require a large physical structure. Our group has recently reported an UCS protocol in zebrafish with several advantages compared to rodent models. We observed that UCS induced behavioral, biochemical, and molecular changes similar to those observed in depressed patients, supporting the translational relevance of the protocol.ObjectivesConsidering that a pharmacological assessment is lacking in this zebrafish model, our aim was to evaluate the effects of anxiolytic (bromazepam) and antidepressant drugs (fluoxetine and nortriptyline) on behavioral (novel tank test), biochemical (whole-body cortisol), and molecular parameters (cox-2, tnf-α, il-6, and il-10 gene expression) in zebrafish subjected to UCS.ResultsWe replicated previous data showing that UCS induces behavioral and neuroendocrine alterations in zebrafish, and we show for the first time that anxiolytic and antidepressant drugs are able to prevent such effects. Furthermore, we extended the molecular characterization of the model, revealing that UCS increases expression of the pro-inflammatory markers cox-2 and il-6, which was also prevented by the drugs tested.ConclusionsThis study reinforces the use of zebrafish as a model organism to study the behavioral and physiological effects of stress. The UCS protocol may also serve as a screening tool for evaluating new drugs that can be used to treat psychiatric disorders with stress-related etiologies.

Astaxanthin prevents changes in the activities of thioredoxin reductase and paraoxonase in hypercholesterolemic rabbits

This study explored the effects of the antioxidant astaxanthin on paraoxonase and thioredoxin reductase activities as well as on other oxidative stress parameters and on the lipid profile in hypercholesterolemic rabbits. Rabbits were fed a standard or a hypercholesterolemic diet alone or supplemented with 50, 100 and 500 mg/100 g of astaxanthin for 60 days. Antioxidant enzymes activities, lipid profile and oxidative stress markers were evaluated in the serum. The hypercholesterolemic diet increased lipids, including unsaturated fatty acids level, whereas it decreased saturated fatty acids level. These changes were accompanied by increased levels of oxidized low-density lipoprotein and oxidized low-density lipoprotein antibodies, as well as lipid and protein oxidation. Astaxanthin (100 and 500 mg/100 g) prevented hypercholesterolemia-induced protein oxidation, whereas 500 mg/100 g of astaxanthin decreased protein oxidation per se. The activities of superoxide dismutase and thioredoxin reductase were enhanced, whereas paraoxonase activity was inhibited in hypercholesterolemic rabbits. All astaxanthin doses prevented changes in thioredoxin reductase and paraoxonase activities. This effect was not related to a direct effect of astaxanthin on these enzymes, because in vitro astaxanthin enhanced thioredoxin reductase and had no effect on paraoxonase activity. Astaxanthin could be helpful in cardiovascular diseases by restoring thioredoxin reductase and paraoxonase activities.

Subchronic atrazine exposure changes defensive behaviour profile and disrupts brain acetylcholinesterase activity of zebrafish

Animal behaviour is the interaction between environment and an individual organism, which also can be influenced by its neighbours. Variations in environmental conditions, as those caused by contaminants, may lead to neurochemical impairments altering the pattern of the behavioural repertoire of the species. Atrazine (ATZ) is an herbicide widely used in agriculture that is frequently detected in surface water, affecting non-target species. The zebrafish is a valuable model organism to assess behavioural and neurochemical effects of different contaminants since it presents a robust behavioural repertoire and also all major neurotransmitter systems described for mammalian species. The goal of this study was to evaluate the effects of subchronic ATZ exposure in defensive behaviours of zebrafish (shoaling, thigmotaxis, and depth preference) using the split depth tank. Furthermore, to investigate a putative role of cholinergic signalling on ATZ-mediated effects, we tested whether this herbicide alters acetylcholinesterase (AChE) activity in brain and muscle preparations. Fish were exposed to ATZ for 14days and the following groups were tested: control (0.2% acetone) and ATZ (10 and 1000μg/L). The behaviour of four animals in the same tank was recorded for 6min and biological samples were prepared. Our results showed that 1000μg/L ATZ significantly increased the inter-fish distance, as well as the nearest and farthest neighbour distances. This group also presented an increase in the shoal area with decreased social interaction. No significant differences were detected for the number of animals in the shallow area, latency to enter the shallow and time spent in shallow and deep areas of the apparatus, but the ATZ 1000 group spent significantly more time near the walls. Although ATZ did not affect muscular AChE, it significantly reduced AChE activity in brain. Exposure to 10μg/L ATZ did not affect behaviour or AChE activity. These data suggest that ATZ impairs defensive behaviours of zebrafish, which could be related to its action on brain cholinergic neurotransmission. Moreover, the use of the split depth tank could be an alternative strategy to assess group behaviour and depth preference after exposure to chemical compounds.

Acute restraint stress induces an imbalance in the oxidative status of the zebrafish brain.

The zebrafish (Danio rerio) has become an emergent model organism for translational approaches focused on the neurobiology of stress due to its genetic, neuroanatomical, and histological similarities with mammalian systems. However, despite the increasing number of studies using zebrafish, reports examining the impact of stress on relevant neurochemical parameters are still elementary when compared to studies using rodents. Additionally, it is important to further validate this model organism by comparing its stress response with those described in other species. Here, we evaluated the effects of an acute restraint stress (ARS) protocol on oxidative stress-related parameters in the zebrafish brain. Our data revealed that ARS significantly decreased catalase activity without altering the activity of superoxide dismutase. Oxidative stress was also indicated by increased levels of lipid peroxides. ARS significantly increased the levels of non-protein thiols, although significant changes in total reduced sulfhydryl content were not detected. These results suggest that ARS is an interesting strategy for evaluating the mechanisms underlying the neurochemical basis of the oxidative profile triggered by acute stressors in the zebrafish brain. Furthermore, this protocol may be suitable for screening new compounds with protective properties against oxidative stress, which plays an increasingly important role in many psychiatric disorders.

Imbalance in superoxide dismutase/thioredoxin reductase activities in hypercholesterolemic subjects: relationship with low density lipoprotein oxidation

BackgroundThere is a relationship among hypercholesterolemia, oxidative stress and inflammation in the atherogenesis. Thus, the objective of the present study was to assess paraoxonase (PON1), superoxide dismutase (SOD) and thioredoxin reductase (TrxR-1) activities and their relationship with lipids, oxidative stress and inflammation in subjects with different low density lipoprotein-cholesterol (LDL) levels.MethodsSerum lipids, highly sensitive C-reactive protein (hs-CRP), lipid and protein oxidation, oxidized LDL (LDLox) and LDLox autoantibodies (LDLoxAB) levels and enzymes activities were measured in a total of 116 subjects that were divided into the following groups according to their LDL levels: low-LDL group (LDL < 100 mg/dL, n = 23), intermediate-LDL group (LDL 100–160 mg/dL, n = 50) and high-LDL group (LDL > 160 mg/dL, n = 43).ResultsThe LDLox and hs-CRP levels increased in the high-LDL group (2.7- and 3.7- fold, respectively), whereas the intermediate and high-LDL groups had higher LDLoxAB (2.2- and 3.1-fold) when compared to low-LDL group (p < 0.05). Similarly, SOD activity, the atherogenic index (AI) and protein oxidation were also higher in the intermediate (1.3-, 1.3- and 1.2-fold) and high-LDL (1.6-, 2.3- and 1.6-fold) groups when compared to the low-LDL group (p < 0.05). Lipid oxidation and SOD/TrxR-1 ratio increased only in the high-LDL group (1.3- and 1.6-fold) when compared to the low-LDL group (p < 0.05). The SOD/TrxR-1 ratio was positively correlated to TBARS (r = 0.23, p < 0.05), LDLox (r = 0.18, p < 0.05), LDLoxAB (r = 0.21, p < 0.05), LDL (r = 0.19, p < 0.05) and AI (r = 0.22, p < 0.05). PON1 and TrxR-1 activities were similar among groups.ConclusionsSome oxidative events initiate when LDL levels are clinically acceptable. Moreover, hypercholesterolemic patients have an imbalance in SOD and TrxR-1 activities that is positively associated to LDL oxidation.

Lead content of dietary calcium supplements available in Brazil

The lead and calcium content of calcium supplements available in Brazil were determined by graphite furnace and flame atomic absorption spectrometry, respectively. Samples were microwave-digested in concentrated HNO3. Citric acid was used as a chemical modifier in the lead analysis. Supplements were classified into six categories: oyster industrialized (OI, n = 4), oyster prepared in pharmacy (OP, n = 3), refined industrialized (RI, n = 6), refined prepared in pharmacy (RP, n = 3), bone meal (B, n = 3), and dolomite (D, n = 4). Lead levels (µg g−1 of measured calcium) were higher in D products (2.33), followed by OI, RP, OP, and RI products (1.46, 1.32, 1.29, 0.75), while B products had levels lower than the limit of quantification (0.02 µg g−1 unit weight). Daily lead intake of eight supplements exceeded the limit of California, USA (1.5 µg g−1 calcium), but none exceeded the federal limit of USA (7.5 µg g−1 calcium) or the provisional tolerable lead intake by FAO/WHO (25 µg kg−1 per week).

Hypolipidemic effects of Solidago chilensis hydroalcoholic extract and its major isolated constituent quercetrin in cholesterol-fed rats

Abstract Context: Despite several studies on the effects of Solidago chilensis Meyen (Asteraceae), the phytochemical and hypolipidemic properties remain underappreciated. Objective: This study evaluates the hypolipidemic and antioxidant effects of hydroalcoholic extract (HE) and quercetrin from S. chilensis aerial parts in cholesterol-fed rats. Materials and methods: The HE was analyzed by high-performance liquid chromatography, followed by quercetrin isolation. Hypercholesterolemic rats (1% cholesterol and 0.5% cholic acid for 15 d) were treated with HE (150, 300, and 600 mg/kg p.o.; n = 6), simvastatin (4 mg/kg p.o.; n = 6), or quercetrin (10 mg/kg p.o.; n = 6) once a day for 30 d. During this period, a high-cholesterol diet was maintained until the 30th day of treatment. Results: Rats treated with HE (150, 300, and 600 mg/kg) and quercetrin showed decreased serum levels of total cholesterol (−19.9, −27.5, −31.0, and −39.4%), lipoprotein-cholesterol (−36.0, −37.5, −43.3, and −59.4%), and triacylglycerides (−15.6, −23.5, −29.8, and −27.2%) when compared with the control group similar to simvastatin. Moreover, treatment with HE and quercetrin decreased hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity (35.1% on average) and increased fecal cholesterol levels (38.2% on average). Discussion and conclusions: Our results suggest that hypolipidemic effects of HE are associated with it modulating the activity of HMG-CoA reductase and its interference in the reabsorption and/or excretion of intestinal lipids. Solidago chilensis and its main constituent, quercetrin, may thus be effective as cholesterol-lowering agents and in preventing atherosclerosis.

Antidepressant-Like and Antioxidant Effects of Plinia trunciflora in Mice

The jaboticaba tree, Plinia trunciflora (O. Berg) Kausel, is popularly named “jabuticabeira” in Brazil and is used in folk medicine to treat diabetes and chronic inflammation of the tonsils, but studies evaluating the central effects of this species are limited. This study evaluated the antidepressant-like and antioxidant effects of P. trunciflora (PT) aqueous extract, in which five different anthocyanins were identified. PT showed significant ferric-reduction power and DPPH radical scavenging activity in vitro and reduced lipid peroxidation both in vitro and ex vivo. At the behavioural level, PT (400 and 800 mg/kg, i.p.) dose-dependently reduced immobility time in the tail suspension test in Swiss male mice. The identification of bioactive compounds accompanied by the in vitro and ex vivo antioxidant activity of PT suggests that these activities might be related to the antidepressant-like activity of P. trunciflora.

Involvement of the catecholaminergic system on the antidepressant-like effects of Alpinia zerumbet in mice

Abstract Context: The traditional uses of Alpinia zerumbet (Pers.) B.L.Burtt & R.m.SM (Zingiberaceae), popularly known as colonia or pacová, suggest that the species has antihypertensive, diuretic, and sedative properties. We previously reported that an ethanol extract of Alpinia zerumbet (HEA) significantly reduced the immobility time in the tail suspension test (TST), similar to the tricyclic antidepressant imipramine. Moreover, HEA presented antioxidant and anxiolytic-like effects in mice. Objective: The objective of this study is to investigate the involvement of monoaminergic and glutamatergic systems in the antidepressant-like effects of this species. Materials and methods: A hydroethanolic extract prepared with the leaves of A. zerumbet was assayed in the TST in male Swiss mice (800 mg/kg, p.o.). Synthesis inhibitors (AMPT, inhibitor of tyrosine hydroxylase, 100 mg/kg, i.p.; and PCPA, irreversible tryptophan hydroxylase inhibitor, 100 mg/kg, i.p.) and a specific glutamate antagonist (AMPA receptor antagonist NBQX, 10 mg/kg, i.p.) were used prior testing. Results: Pre-treatment with the noradrenergic/dopaminergic inhibitor AMPT fully abolished the anti-immobility effects of HEA, with the two-way ANOVA yielding a significant interaction between pre-treatment and treatment (F1,32 = 10.0, p < 0.01); no interaction was observed with the serotonergic inhibitor PCPA (F1,32 = 0.33, p > 0.05) or NBQX (F1,32 = 0.21, p > 0.05). Conclusion: These results indicated that HEA most likely acts through the dopaminergic and/or noradrenergic system but not through the serotoninergic or glutamatergic systems. This study reinforces the idea that the available biodiversity in Brazil can serve as a basis for innovation in the development of new drugs.