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Background: Sepsis is a severe global health problem, with high morbidity and mortality. In sepsis, one of the main affected organs is the liver. Hepatic alterations characterize a negative prognostic. Omega-3 fatty acids (ω3), eicosapentaenoic acid, and docosahexaenoic acid, are part of the main families of polyunsaturated fatty acids. ω3 has been used in studies as sepsis treatment and as a treatment for non-alcoholic liver disease. Aim: We aimed to evaluate the effects of treatment with fish oil (FO) rich in ω3 on liver changes and damage resulting from experimental sepsis. Methodology: A model of severe sepsis in Wistar rats was used. Oxidative stress in the liver tissue was evaluated by means of tests of thiobarbituric acid reactive substances, 2,7-dihydrodichlorofluorescein diacetate , catalase, and glutathione peroxidase, in the serum TBARS, DCF, thiols and, to assess liver dysfunction, alanine aminotransferase and aspartate aminotransferase. Hepatic tissue damage was evaluated using H&E histology. Results: In assessments of oxidative stress in liver tissue, a protective effect was observed in the tests of TBARS, DCF, CAT, and GPx, when compared the sepsis versus sepsis+ω3 groups. Regarding the oxidative stress in serum, a protective effect of treatment with ω3 was observed in the TBARS, DCF, and thiols assays, in the comparison between the sepsis and sepsis+ω3 groups. ω3 had also a beneficial effect on biochemical parameters in serum in the analysis of ALT, creatinine, urea, and lactate, observed in the comparison between the sepsis and sepsis+ω3 groups. Conclusion: The results suggest ω3 as a liver protector during sepsis with an antioxidant effect, alleviating injuries and dysfunctions.
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Reactive oxygen species and reactive nitrogen species (RONS) are involved in programmed cell death in the context of numerous degenerative and chronic diseases. In particular, the ability of cells to maintain redox homeostasis is necessary for an adaptive cellular response to adverse conditions that can cause damage to proteins and DNA, resulting in apoptosis and genetic mutations. Here, we focus on the 2',7'-dichlorodihydrofluorescein diacetate (DCFH2-DA) assay to detect RONS. Although this fluorescence-based assay is widely utilized due to its high sensitivity to detect changes in cellular redox status that allow measuring alterations in RONS over time, its validity has been a matter of controversy. If correctly carried out, its limitations are understood and results are correctly interpreted, the DCFH2-DA assay is a valuable tool for cell-based studies.
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BACKGROUND: Deoxymikanolide is a sesquiterpene lactone isolated from Mikania micrantha and M. variifolia which, has previously demonstrated in vitro activity on Trypanosoma cruzi and in vivo activity on an infected mouse model. PURPOSE: Based on these promising findings, the aim of this study was to investigate the mechanism of action of this compound on different parasite targets. METHODS: The interaction of deoxymikanolide with hemin was examined under reducing and non- reducing conditions by measuring modifications in the Soret absorption band of hemin; the thiol interaction was determined spectrophotometrically through its reaction with 5,5'-dithiobis-2-nitrobenzoate in the presence of glutathione; activity on the parasite antioxidant system was evaluated by measuring the activity of the superoxide dismutase and trypanothione reductase enzymes, together with the intracellular oxidative state by flow cytometry. Superoxide dismutase and trypanothione reductase activities were spectrophotometrically tested. Cell viability, phosphatidylserine exposure and mitochondrial membrane potential were assessed by means of propidium iodide, annexin-V and rhodamine 123 staining, respectively; sterols were qualitatively and quantitatively tested by TLC; ultrastructural changes were analyzed by transmission electron microscopy. Autophagic cells were detected by staining with monodansylcadaverine. RESULTS: Deoxymikanolide decreased the number of reduced thiol groups within the parasites, which led to their subsequent vulnerability to oxidative stress. Treatment of the parasites with the compound produced a depolarization of the mitochondrial membrane even though the plasma membrane permeabilization was not affected. Deoxymikanolide did not affect the intracellular redox state and so the mitochondrial dysfunction produced by this compound could not be attributed to ROS generation. The antioxidant defense system was affected by deoxymikanolide at twenty four hours of treatment, when both an increased oxidative stress and decreased activity of superoxide dismutase and trypanothione reductase (40 and 60% respectively) were observed. Both the oxidative stress and mitochondrial dysfunction induce parasite death by apoptosis and autophagy. CONCLUSION: Based on our results, deoxymikanolide would exert its anti-T cruzi activity as a strong thiol blocking agent and by producing mitochondrial dysfunction.
Assuntos
Lactonas/farmacologia , Sesquiterpenos de Germacrano/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Glutationa/metabolismo , Hemina/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mikania/química , NADH NADPH Oxirredutases/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Esteróis/biossíntese , Superóxido Dismutase/metabolismo , Trypanosoma cruzi/patogenicidade , Trypanosoma cruzi/ultraestruturaRESUMO
A Displasia Coxofemoral (DCF) em cães é uma afecção definida como uma doença multifatorial, determinada por fatores genéticos, nutricionais, muito influenciada pelo ambiente local de domesticação (Rocha,2018). A DCF compromete a biomecânica e representa a disparidade entre a massa muscular e o rápido crescimento ósseo, levando uma instabilidade na articulação do quadril (Fossum,2007). Este trabalho tem como objetivo relatar a melhora de um canino com DCF grave submetido ao tratamento por ozonioterapia e nutracêuticos.
Assuntos
Animais , Cães , Displasia Pélvica Canina/terapia , Displasia Pélvica Canina/tratamento farmacológico , Ozônio/uso terapêutico , Suplementos Nutricionais , Coxeadura AnimalRESUMO
Foi atendido um cão de dois anos, SRD, com diagnostico de shunt portossistêmico em uma clínica particular de São José dos Campos. O mesmo deu entrada para atendimento clinico com hematuria e disúria. Ao exame de urina e de ultrassonografia (US), constatou-se uma formação de urólito na bexiga com as dimensões de 1,15 x 0,52 cm. Tendo em vista a dificuldade de manejo clínico de urolitos em cães, a fim de se evitar o procedimento cirúrgico o mesmo foi encaminhado para ozonioterapia, por ser um tratamento menos invasivo para um animal jovem, porém com uma doença de fundo congênito devido à ocorrência ter predisposições raciais entre outras causas.
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Animais , Cães , Ozônio/uso terapêutico , Urolitíase/terapia , Urolitíase/veterinária , Cálculos Urinários/terapia , Cálculos Urinários/veterináriaRESUMO
Foi atendido um cão de dois anos, SRD, com diagnostico de shunt portossistêmico em uma clínica particular de São José dos Campos. O mesmo deu entrada para atendimento clinico com hematuria e disúria. Ao exame de urina e de ultrassonografia (US), constatou-se uma formação de urólito na bexiga com as dimensões de 1,15 x 0,52 cm. Tendo em vista a dificuldade de manejo clínico de urolitos em cães, a fim de se evitar o procedimento cirúrgico o mesmo foi encaminhado para ozonioterapia, por ser um tratamento menos invasivo para um animal jovem, porém com uma doença de fundo congênito devido à ocorrência ter predisposições raciais entre outras causas.(AU)
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Animais , Cães , Ozônio/uso terapêutico , Urolitíase/terapia , Urolitíase/veterinária , Cálculos Urinários/terapia , Cálculos Urinários/veterináriaRESUMO
A Displasia Coxofemoral (DCF) em cães é uma afecção definida como uma doença multifatorial, determinada por fatores genéticos, nutricionais, muito influenciada pelo ambiente local de domesticação (Rocha,2018). A DCF compromete a biomecânica e representa a disparidade entre a massa muscular e o rápido crescimento ósseo, levando uma instabilidade na articulação do quadril (Fossum,2007). Este trabalho tem como objetivo relatar a melhora de um canino com DCF grave submetido ao tratamento por ozonioterapia e nutracêuticos.(AU)
Assuntos
Animais , Cães , Displasia Pélvica Canina/tratamento farmacológico , Displasia Pélvica Canina/terapia , Suplementos Nutricionais , Ozônio/uso terapêutico , Coxeadura AnimalRESUMO
Parkinson's disease (PD) is characterized by progressive dopamine (DA) depletion in the striatum. Exercise has been shown to be a promising non-pharmacological approach to reduce the risk of neurodegeneration diseases. This study was designed to investigate the potential neuroprotective effect of swimming training (ST) in a mouse model of PD induced by 6-hydroxydopamine (6-OHDA) in mice. The present study demonstrated that a 4-week ST was effective in attenuating the following impairments resulting from 6-OHDA exposure: (i) depressive-like behavior in the tail suspension test; (ii) increase in the number of falls in the rotarod test; (iii) impairment on long-term memory in the object recognition test; (iv) increase of the reactive species and interleukin 1-beta (IL-1ß) levels; (v) inhibition of the glutathione peroxidase (GPx) activity; (vi) rise of the glutathione reductase (GR) and glutathione S-transferase (GST) activities and vii) decrease of DA, homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) levels. The mechanisms involved in this study are the modulation of GPx, GR and GST activities as well as IL-1ß level in a PD model induced by 6-OHDA, protecting against the decrease of DA, DOPAC and HVA levels in the striatum of mice. These findings reinforce that one of the effects induced by exercise on neurodegenerative disease, such as PD, is due to antioxidant and anti-inflammatory properties. We suggest that exercise attenuates cognitive and motor declines, depression, oxidative stress, and neuroinflammation induced by 6-OHDA supporting the hypothesis that exercise can be used as a non-pharmacological tool to reduce the symptoms of PD.
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Terapia por Exercício/métodos , Hidroxidopaminas/toxicidade , Doença de Parkinson/etiologia , Doença de Parkinson/reabilitação , Natação/fisiologia , Animais , Catalase/metabolismo , Citrato (si)-Sintase/metabolismo , Corpo Estriado/enzimologia , Depressão/etiologia , Modelos Animais de Doenças , Comportamento Exploratório/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Elevação dos Membros Posteriores , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/enzimologia , Doença de Parkinson/fisiopatologia , Desempenho Psicomotor , Reconhecimento Psicológico , Teste de Desempenho do Rota-RodRESUMO
It is suggested that systemic oxidative stress and inflammation play a central role in the onset and progression of cardiovascular diseases associated with the exposure to particulate matter (PM). The aim of this work was to evaluate the time changes of systemic markers of oxidative stress and inflammation, after an acute exposure to Residual Oil Fly Ash (ROFA). Female Swiss mice were intranasally instilled with a ROFA suspension (1.0mg/kg body weight) or saline solution, and plasma levels of oxidative damage markers [thiobarbituric acid reactive substances (TBARSs) and protein carbonyls], antioxidant status [reduced (GSH) and oxidized (GSSG) glutathione, ascorbic acid levels, and superoxide dismutase (SOD) activity], cytokines levels, and intravascular leukocyte activation were evaluated after 1, 3 or 5h of exposure. Oxidative damage to lipids and decreased GSH/GSSG ratio were observed in ROFA-exposed mice as early as 1h. Afterwards, increased protein oxidation, decreased ascorbic acid content and SOD activity were found in this group at 3h. The onset of an adaptive response was observed at 5h after the ROFA exposure, as indicated by decreased TBARS plasma content and increased SOD activity. The observed increase in oxidative damage to plasma macromolecules, together with systemic antioxidants depletion, may be a consequence of a systemic inflammatory response triggered by the ROFA exposure, since increased TNF-α and IL-6 plasma levels and polymorphonuclear leukocytes activation was found at every evaluated time point. These findings contribute to the understanding of the increase in cardiovascular morbidity and mortality, in association with environmental PM inhalation.
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Cinza de Carvão/toxicidade , Inflamação/patologia , Estresse Oxidativo/efeitos dos fármacos , Administração por Inalação , Animais , Antioxidantes/metabolismo , Ácido Ascórbico/sangue , Feminino , Glutationa/sangue , Coração/efeitos dos fármacos , Coração/fisiopatologia , Inflamação/induzido quimicamente , Interleucina-6/sangue , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Camundongos , Neutrófilos/citologia , Neutrófilos/metabolismo , Carbonilação Proteica/efeitos dos fármacos , Superóxido Dismutase/sangue , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fator de Necrose Tumoral alfa/sangueRESUMO
Sulfite oxidase (SO) deficiency is biochemically characterized by tissue accumulation and high urinary excretion of sulfite, thiosulfate and S-sulfocysteine. Affected patients present severe neurological symptoms and cortical atrophy, whose pathophysiology is still poorly established. Therefore, in the present work we investigated the in vitro effects of sulfite and thiosulfate on important parameters of energy metabolism in the brain of young rats. We verified that sulfite moderately inhibited the activity of complex IV, whereas thiosulfate did not alter any of the activities of the respiratory chain complexes. It was also found that sulfite and thiosulfate markedly reduced the activity of total creatine kinase (CK) and its mitochondrial and cytosolic isoforms, suggesting that these metabolites impair brain cellular energy buffering and transfer. In contrast, the activity of synaptic Na(+),K(+)-ATPase was not altered by sulfite or thiosulfate. We also observed that the inhibitory effect of sulfite and thiosulfate on CK activity was prevented by melatonin, reduced glutathione and the combination of both antioxidants, as well as by the nitric oxide synthase N(ω)-nitro-l-arginine methyl ester, indicating the involvement of reactive oxygen and nitrogen species in these effects. Sulfite and thiosulfate also increased 2',7'-dichlorofluorescin oxidation and hydrogen peroxide production and decreased the activity of the redox sensor aconitase enzyme, reinforcing a role for oxidative damage in the effects elicited by these metabolites. It may be presumed that the disturbance of cellular energy and redox homeostasis provoked by sulfite and thiosulfate contributes to the neurological symptoms and abnormalities found in patients affected by SO deficiency.
Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/complicações , Encefalopatias Metabólicas/etiologia , Encéfalo/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Sulfito Oxidase/deficiência , Sulfitos/farmacologia , Tiossulfatos/farmacologia , Erros Inatos do Metabolismo dos Aminoácidos/genética , Erros Inatos do Metabolismo dos Aminoácidos/metabolismo , Erros Inatos do Metabolismo dos Aminoácidos/fisiopatologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Encéfalo/fisiologia , Encefalopatias Metabólicas/genética , Encefalopatias Metabólicas/metabolismo , Transporte de Elétrons/efeitos dos fármacos , Transporte de Elétrons/genética , Transporte de Elétrons/fisiologia , Metabolismo Energético/fisiologia , Masculino , Oxirredução/efeitos dos fármacos , Ratos , Ratos Wistar , Sulfito Oxidase/genética , Sulfito Oxidase/metabolismo , Sulfitos/metabolismo , Tiossulfatos/metabolismoRESUMO
Novel ß-lapachone analogs 2-phenyl-3,4-dihydro-2H-benzo[h]chromene-5,6-dione (NQ1), 2-p-tolyl-3,4-dihydro-2H-benzo[h]chromene-5,6-dione (NQ3) and 2-methyl-2-phenyl-3,4-dihydro-2H-benzo[h]chromene-5,6-dione (NQ7), which have trypanocidal activity, were assayed for cytotoxic effects on murine EL-4 T lymphoma cells. The NQs inhibited the proliferation of EL-4 cells at concentrations above 1µM. Nuclear staining of the EL-4 cells revealed chromatin condensation and a nuclear morphology compatible with the induction of apoptosis. Flow cytometry assays with annexin V-FITC and propidium iodide confirmed the cell death by apoptosis. Using electron paramagnetic resonance (EPR), a semiquinone radical was detected in EL-4 cells treated with NQs. In addition, a decrease in the GSH level in parallel with reactive oxygen species (ROS) production was observed. Preincubation with n-acetyl-l-cysteine (NAC) was able to reverse the inhibitory effects of the NQs on cell proliferation, indicating that ROS generation is involved in NQ-induced apoptosis. In addition, the NQs induced a decrease in the mitochondrial membrane potential and increased the proteolytic activation of caspases 9 and 3 and the cleavage of Poly (ADP-Ribose) Polymerase (PARP). In conclusion, these results indicate that redox cycling is induced by the NQs in the EL-4 cell line, with the generation of ROS and other free radicals that could inhibit cellular proliferation as a result of the induction of the intrinsic apoptosis pathway.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Linfoma de Células T/tratamento farmacológico , Naftoquinonas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima/efeitos dos fármacos , Acetilcisteína/farmacologia , Animais , Antineoplásicos/antagonistas & inibidores , Benzopiranos/antagonistas & inibidores , Benzopiranos/farmacologia , Benzoquinonas/metabolismo , Linhagem Celular Tumoral , Forma do Núcleo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Montagem e Desmontagem da Cromatina/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Glutationa/antagonistas & inibidores , Glutationa/metabolismo , Cinética , Linfoma de Células T/metabolismo , Linfoma de Células T/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos , Naftoquinonas/antagonistas & inibidores , Espécies Reativas de Oxigênio/antagonistas & inibidores , Tripanossomicidas/antagonistas & inibidores , Tripanossomicidas/farmacologiaRESUMO
BACKGROUND: Activation of ATP-gated P2X7 receptors (P2X7R) in macrophages leads to production of reactive oxygen species (ROS) by a mechanism that is partially characterized. Here we used J774 cells to identify the signaling cascade that couples ROS production to receptor stimulation. METHODS: J774 cells and mP2X7-transfected HEK293 cells were stimulated with Bz-ATP in the presence and absence of extracellular calcium. Protein inhibitors were used to evaluate the physiological role of various kinases in ROS production. In addition, phospho-antibodies against ERK1/2 and Pyk2 were used to determine activation of these two kinases. RESULTS: ROS generation in either J774 or HEK293 cells (expressing P2X7, NOX2, Rac1, p47phox and p67phox) was strictly dependent on calcium entry via P2X7R. Stimulation of P2X7R activated Pyk2 but not calmodulin. Inhibitors of MEK1/2 and c-Src abolished ERK1/2 activation and ROS production but inhibitors of PI3K and p38 MAPK had no effect on ROS generation. PKC inhibitors abolished ERK1/2 activation but barely reduced the amount of ROS produced by Bz-ATP. In agreement, the amount of ROS produced by PMA was about half of that produced by Bz-ATP. CONCLUSIONS: Purinergic stimulation resulted in calcium entry via P2X7R and subsequent activation of the PKC/c-Src/Pyk2/ERK1/2 pathway to produce ROS. This signaling mechanism did not require PI3K, p38 MAPK or calmodulin. GENERAL SIGNIFICANCE: ROS is generated in order to kill invading pathogens, thus elucidating the mechanism of ROS production in macrophages and other immune cells allow us to understand how our body copes with microbial infections.
Assuntos
Quinase 2 de Adesão Focal/metabolismo , Sistema de Sinalização das MAP Quinases , Macrófagos/metabolismo , Estresse Oxidativo , Proteínas Proto-Oncogênicas pp60(c-src)/metabolismo , Receptores Purinérgicos P2X7/fisiologia , Animais , Cálcio/metabolismo , Linhagem Celular , Humanos , Transporte de Íons , Macrófagos/enzimologia , CamundongosRESUMO
In order to clarify the role of 2,6-dichlorophenol (2,6-DCP) in the courtship of Amblyomma cajennense, sexually mature males that had previously fed on rabbits were tested in bioassays. The males were released onto dummies treated with whole female extract or synthetic 2,6-DCP at a concentration of two female equivalents, or with hexane (control), and their responses were observed. In the presence of both the extract and 2,6-DCP, excitation was observed among the males, expressed in the form of touching and probing the dummy, and mounting occurred readily. The percentages of mounting (73 percent) and tipping over (60 percent) were equal in the two treatments and higher than in the control group (27 and 20 percent, respectively). Relatively short durations of mounting were recorded, and these were statistically similar in all treatments. Almost all instances of mounting resulted in tipping-over behavior. A few isolated cases of males that went directly to ventral positioning without mounting were observed. It was confirmed that 2,6-DCP alone is capable of mediation of mounting behavior in A. cajennense.
Visando elucidar o papel do 2,6-diclorofenol (2,6-DCF) no cortejo de Amblyomma cajennense, machos sexualmente maduros, previamente alimentados em coelhos, foram avaliados em testes biológicos. Os machos foram liberados sobre manequins tratados com um extrato de fêmeas, ou com 2,6-DCF sintético na concentração equivalente a duas fêmeas, ou com hexano (controle), e suas respostas foram observadas. Na presença do extrato e do 2,6-DCF, a excitação dos machos foi expressa na forma de toques e sondagens, e a monta ocorreu rapidamente. As porcentagens de respostas observadas nos dois tratamentos foram iguais, sendo a monta (73 por cento) e retorno na superfície ventral (60 por cento) mais altos que no controle (27 e 20 por cento, respectivamente). Foram observados períodos de monta relativamente curtos, sendo esses estatisticamente iguais em todos os tratamentos, e quase todos resultando em posicionamento ventral. Alguns casos isolados de posicionamento ventral sem monta foram observados. Foi confirmado que o 2,6-DCP sozinho é capaz de mediar o comportamento de monta de A. cajennense.