Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 285
Filtrar
1.
Physiol Int ; 106(2): 158-167, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31271310

RESUMO

Obesity is related to increased oxidative stress. Although low-intensity physical exercise reduces oxidative stress, obese subjects may show exercise intolerance. For these subjects, inspiratory threshold loading could be an alternative tool to reduce oxidative stress. We investigated the effects of inspiratory threshold loading on biomarkers of oxidative stress in obese and normal-weight subjects. Twenty obese (31.4 ± 6 years old, 10 men and 10 women, 37.5 ± 4.7 kg/m2) and 20 normal-weight (29.4 ± 8 years old, 10 men and 10 women, 23.2 ± 1.5 kg/m2) subjects matched for age and gender participated in the study. Maximal inspiratory pressure (MIP) was assessed by a pressure transducer. Blood sampling was performed before and after loading and control protocols to assess thiobarbituric acid reactive substances (TBARS), protein carbonylation, and reduced glutathione. Inspiratory threshold loading was performed at 60% MIP and maintained until task failure. The 30-min control protocol was performed at 0 cmH2O. Our results demonstrated that inspiratory threshold loading reduced TBARS across time in obese (6.21 ± 2.03 to 4.91 ± 2.14 nmol MDA/ml) and normal-weight subjects (5.60 ± 3.58 to 4.69 ± 2.80 nmol MDA/ml; p = 0.007), but no change was observed in protein carbonyls and glutathione in both groups. The control protocol showed no significant changes in TBARS and protein carbonyls. However, reduced glutathione was increased across time in both groups (obese: from 0.50 ± 0.37 to 0.56 ± 0.35 µmol GSH/ml; normal-weight: from 0.61 ± 0.11 to 0.81 ± 0.23 µmol GSH/ml; p = 0.002). These findings suggest that inspiratory threshold loading could be potentially used as an alternative tool to reduce oxidative stress in both normal-weight and obese individuals.


Assuntos
Inalação/fisiologia , Peroxidação de Lipídeos/fisiologia , Obesidade/fisiopatologia , Adulto , Biomarcadores/metabolismo , Exercício Físico/fisiologia , Feminino , Glutationa/metabolismo , Humanos , Masculino , Obesidade/metabolismo , Estresse Oxidativo/fisiologia , Carbonilação Proteica/fisiologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Pesos e Medidas
2.
Res Q Exerc Sport ; 90(3): 385-394, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31135277

RESUMO

Purpose: The purpose of this study was to investigate (a) time-dependent changes in muscle damage (MD) biomarkers, oxidative stress (OS) indices, and maximum strength performance; (b) the relationship between changes in maximum strength performance and changes in MD and OS indices; and (c) whether eccentric exercise-induced MD is related to OS. Method: Twenty-nine male volunteers (age: 22.13 ± 3.1 years) participated in the study. Participants performed 60 maximal eccentric actions of the elbow flexors at a constant velocity of 60°·s-1. Maximum isokinetic strength (MIS), visual analog scale soreness scores, serum creatine kinase (CK) activity, total antioxidant status, total oxidant status (TOS), protein carbonyl (PCO), and 8-hydroxydeoxyguanosine level were analyzed. Blood samples were obtained before, immediately after, and 24 h, 48 h, and 96 h after the eccentric exercise. Change in total work (%ΔTWk), peak torque (%ΔPT), and OS index were calculated. Results: CK, PCO, and TOS significantly increased over time (p < .05). However, no significant main effect was observed for MIS or any other investigated biomarkers (p > .05). MIS was not related to MD or OS indices. However, %ΔTWk demonstrated a moderate inverse correlation with OS indices. No significant relationship was observed between %ΔPT and any of the selected biomarkers. Conclusions: Our findings confirm the hypothesis that acute eccentric exercise increases MD biomarkers and OS indices. However, indices of OS damage were significantly related, particularly, to the strength loss of flexors. This finding suggests that the decline in strength performance is not the primary determinant of the magnitude of MD following voluntary eccentric contraction.


Assuntos
Exercício Físico/fisiologia , Força Muscular/fisiologia , Músculo Esquelético/lesões , Estresse Oxidativo/fisiologia , 8-Hidroxi-2'-Desoxiguanosina/sangue , Adulto , Biomarcadores/sangue , Creatina Quinase/sangue , Articulação do Cotovelo/fisiologia , Humanos , Contração Isométrica/fisiologia , Masculino , Músculo Esquelético/fisiologia , Mialgia/etiologia , Mialgia/fisiopatologia , Carbonilação Proteica/fisiologia , Adulto Jovem
3.
Eur J Dermatol ; 29(S1): 11-14, 2019 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-31017576

RESUMO

Ageing and age-related diseases (ARD) share a common biological clock that appears as their root cause: protein damage. A majority of proteins have evolved into native structures resistant to oxidative damage but any folding imperfections, including those due to "silent" amino acid substitutions, reduce oxidation resistance. Damaged proteins accumulate with age and trigger ageing-like phenotypes reversible by their turnover, while acquired genome alterations remain as stable consequences of protein malfunction. Ageing and ARD display species-specific latency in phenotypic expression. Disease latency may be proposed as to be due to phenotypic suppression of cellular defects by molecular traffic among neighbouring cells. Such cross-complementation of functional deficiencies acts as a kind of tissue-based cellular "solidarity", called cellular parabiosis. Chronic inflammation reveals dormant cell phenotypes and shortens disease latency by the breakdown of cell-cell communication, as in tumour promotion and inflammation. At the present time, predictive diagnostics, prognostics, prevention and even cure of disease by phenotypic reversion become conceivable.


Assuntos
Envelhecimento/fisiologia , Comunicação Celular/fisiologia , Meio Ambiente , Carbonilação Proteica/fisiologia , Fenômenos Fisiológicos Celulares , Dano ao DNA/fisiologia , Ecossistema , Homeostase , Humanos , Estresse Oxidativo/fisiologia , Espécies Reativas de Oxigênio/efeitos adversos
4.
Andrologia ; 51(4): e13233, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30637798

RESUMO

The cryopreservation of sperm is a well established technique that plays an essential role in dissemination of elite germplasm of livestock. Despite having numerous advantages, the cryopreservation induces certain stresses on sperm including structural and functional damages leading to impaired sperm quality and fertility, which might be associated with production of reactive oxygen species (ROS). In addition, the ROS upon reacting with sperm lipids, DNA and proteins may lead to a cascade of sperm damages. The sperm membrane contains a rich amount of polyunsaturated fatty acids, which increases their susceptibility to oxidative stress induced damages, leading to formation of secondary products. These secondary products result in oxidation of sperm proteins via carbonylation. The carbonylation could lead to disturbances in specific proteins that are involved in capacitation. The present review deals with sperm protein carbonylation.


Assuntos
Criopreservação , Congelamento/efeitos adversos , Carbonilação Proteica/fisiologia , Preservação do Sêmen/efeitos adversos , Espermatozoides/metabolismo , Criação de Animais Domésticos/métodos , Animais , Cruzamento/métodos , Masculino , Espécies Reativas de Oxigênio/metabolismo , Preservação do Sêmen/métodos , Capacitação Espermática/fisiologia
5.
Artigo em Inglês | MEDLINE | ID: mdl-30476595

RESUMO

Oxidative damage is a potential physiological cost of thermoregulation during seasonal adjustments to air temperature (Ta) in small mammals. Here, we hypothesized that Ta affects serum thyroid hormone levels and these hormones can mediate the changes in metabolic rate and oxidative damage. Mongolian gerbils (Meriones unguiculatus) were acclimated at different Tas (5 °C, 23 °C and 37 °C) for 3 weeks. Serum tri-iodothyronine (T3) levels increased at 5 °C but decreased at 37 °C compared to the control (23 °C). Protein carbonyls increased in liver at 37 °C compared with control, however, lipid damage (malonaldehyde, MDA) in both serum and liver was unrelated to Ta. After the effects of different Tas on thyroid hormone levels and oxidative damage markers were determined, we further investigate whether thyroid hormones mediated metabolic rate and oxidative damage. Another set of gerbils received 0.0036% L-thyroxin (hyperthyroid), 0.04% Methylimazol (hypothyroid) or water (control). Hypothyroid group showed a 34% reduction in resting metabolic rate (RMR) also 42% and 26% increases in MDA and liver protein carbonyl respectively, whereas hyperthyroid group had higher RMR, liver mass and superoxide dismutase (SOD) compared to control. Serum T3 or T3/T4 levels were correlated positively with RMR, liver mass, and SOD, but negatively with MDA and uncoupling protein 2 (UCP2). We concluded that high Ta induced hypothyroidism, decreased RMR and increased oxidative damage, whereas low Ta induced hyperthyroidism, increased RMR and unchanged oxidative damage. These data supported our hypothesis that thyroid hormones can be a cue to mediate metabolic rate and different aspects of oxidative and antioxidant activities at different Tas.


Assuntos
Metabolismo Basal/fisiologia , Gerbillinae/fisiologia , Oxirredução , Hormônios Tireóideos/fisiologia , Animais , Antioxidantes/metabolismo , Gerbillinae/metabolismo , Fígado/metabolismo , Malondialdeído/metabolismo , Carbonilação Proteica/fisiologia , Superóxido Dismutase/metabolismo , Temperatura , Proteína Desacopladora 2/metabolismo
6.
Oxid Med Cell Longev ; 2018: 5286785, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30581533

RESUMO

Wound healing is a complex multiphase process which can be hampered by many factors including impaired local circulation, hypoxia, infection, malnutrition, immunosuppression, and metabolic dysregulation in diabetes. Redox dysregulation is a common feature of many skin diseases demonstrated by virtually all cell types in the skin with overproduction of reactive oxygen and nitrogen species. The objective of this study was to characterize the redox environment in wound fluids and sera from patients suffering from chronic leg ulcers (n = 19) and acute wounds (bulla fluids from second degree burns; n = 11) with serum data also compared to those from healthy volunteers (n = 7). Significantly higher concentrations of TNF-α, interleukine-8, vascular endothelial growth factor, and lactate dehydrogenase (measure of cell damage) were found in fluids from chronic wounds compared to acute ones. The extent of protein carbonylation (measure of protein oxidation), lipid peroxidation, and tyrosine nitration (indicator of peroxynitrite production) was similar in acute and chronic wound fluids, while radical scavenging activity and glutathione (GSH) levels were elevated in chronic wound fluids compared to acute wounds. Sera were also assessed for the same set of parameters with no significant differences detected. Nitrotyrosine (the footprint of the potent oxidant peroxynitrite) and poly(ADP-ribose) (the product of the DNA damage sensor enzyme PARP-1) could be detected in wound biopsies. Our data identify multiple signs of redox stress in chronic wounds with notable differences. In chronic wounds, elevations in antioxidant levels/activities may indicate compensatory mechanisms against inflammation. The presence of nitrotyrosine and poly(ADP-ribose) in tissues from venous leg ulcers indicate peroxynitrite production and PARP activation in chronic wounds.


Assuntos
Cicatrização/fisiologia , Idoso , Idoso de 80 Anos ou mais , Feminino , Glutationa/metabolismo , Humanos , Interleucina-8/metabolismo , L-Lactato Desidrogenase/metabolismo , Peroxidação de Lipídeos/fisiologia , Masculino , Pessoa de Meia-Idade , Oxirredução , Poli(ADP-Ribose) Polimerase-1/metabolismo , Poli Adenosina Difosfato Ribose/metabolismo , Carbonilação Proteica/fisiologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
7.
Neurochem Res ; 43(12): 2277-2287, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30251207

RESUMO

The present study was designed to investigate the role of calpain and the proteasome in the removal of oxidized neuronal cytoskeletal proteins in myelin basic protein-induced experimental autoimmune encephalomyelitis (EAE). To this end, EAE rats received a single intrathecal injection of calpeptin or epoxomicin at the first sign of clinical disease. Forty-eight hours later, animals were sacrificed and lumbar spinal cord segments were dissected and used for biochemical analyses. The results show that calpain and proteasome activity is specifically, but partially, inhibited with calpeptin and epoxomicin, respectively. Calpain inhibition causes an increase in total protein carbonylation and in the amount of neurofilament proteins (NFPs), ß-tubulin and ß-actin that were spared from degradation, but no changes are seen in the oxidation of any of three NFPs. By contrast, proteasome inhibition has no effect on total protein carbonylation or cytoskeletal protein degradation but increases the amount of oxidized NFH and NFM. These results suggest that while the proteasome may contribute to removal of oxidized NFPs, calpain is the main protease involved in degradation of neuronal cytoskeleton and does not preferentially targets oxidized NFPs species in acute EAE. Different results were obtained in a cell-free system, where calpain inhibition rises the amount of oxidized NFH, and proteasome inhibition fails to change the oxidation state of the NFPs. The later finding suggests that the preferential degradation of oxidized NFH and NFM in vivo by the proteasome occurs via the 26S and not the 20S particle.


Assuntos
Calpaína/fisiologia , Citoesqueleto/metabolismo , Encefalomielite Autoimune Experimental/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Carbonilação Proteica/fisiologia , Proteólise , Animais , Calpaína/antagonistas & inibidores , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/patologia , Dipeptídeos/administração & dosagem , Encefalomielite Autoimune Experimental/patologia , Injeções Espinhais , Masculino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Oligopeptídeos/administração & dosagem , Carbonilação Proteica/efeitos dos fármacos , Proteólise/efeitos dos fármacos , Ratos , Ratos Endogâmicos Lew
8.
J Cell Biochem ; 119(11): 9099-9109, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30076739

RESUMO

Advanced glycation end products (AGEs) are directly related to third aging-associated diseases, such as cardiovascular diseases, arteriosclerosis, and neurodegeneration. Likewise, these irreversible and nonenzymatic products have been reported to be involved in the progression of malignant cancers. In general, aging-associated diseases and the initiation of cancer have been subjects of interest for several years. Few studies on the role of AGEs in cancer have been performed on cell lines. Moreover, past investigations in the field of glycation biology still lack the knowledge of in vivo and in vitro approaches for cancer cells. Accordingly, we aimed to focus on and establish a link between cancer and glycation with respect to all the possible AGEs. In our study, the levels of carboxymethyllysine (CML) increased by 50.94% in an animal model of glycation, whereas in an animal model of cancer, the contents of CML increased by 45.94% compared with their negative controls. Similarly, fluorescent AGEs were also examined and were found to be increased by 65.3% and 58.63% in the animal models of glycation and cancer, respectively, compared with the control subjects. The protein carbonyl contents were also found to be enhanced in the animal models of glycation and cancer. In our study, the levels of reactive oxygen species were also found to be significantly increased in the in vitro model of cancer cells as compared with the controls. Such an initial breakthrough indicated that AGEs were present in the serum of the animal models of cancer and glycation.


Assuntos
Produtos Finais de Glicação Avançada/metabolismo , Neoplasias Pulmonares/metabolismo , Células A549 , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Humanos , Lisina/análogos & derivados , Lisina/metabolismo , Masculino , Camundongos , Carbonilação Proteica/fisiologia , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo
9.
Oxid Med Cell Longev ; 2018: 4149681, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30057679

RESUMO

Patients with end-stage renal disease (ESRD) undergoing haemodialysis (HD) experience oxidative/carbonyl stress, which is postulated to increase after the HD session. The influence of diabetes mellitus and sex on oxidation of plasma proteins in ESRD has not yet been clarified despite that diabetic nephropathy is the most common cause of ESRD in developed and developing countries and despite the increasingly emerging differences between males and females in epidemiology, pathophysiology, clinical manifestations, and outcomes for several diseases. Therefore, this study aimed to evaluate the possible effect of type 2 diabetes mellitus, gender, and dialysis filter on plasma level of protein carbonyls (PCO) in ESRD patients at the beginning and at the end of a single HD session. Results show that mean post-HD plasma PCO levels are significantly higher than mean pre-HD plasma PCO levels and that the type of dialysis filter and dialysis technique are unrelated to plasma PCO levels. The mean level of plasma PCO after a HD session increases slightly but significantly in nondiabetic ESRD patients compared to diabetic ones, whereas it increases more markedly in women than in men. These novel findings suggest that women with ESRD are more susceptible than men to oxidative/carbonyl stress induced by HD.


Assuntos
Diabetes Mellitus Tipo 2/sangue , Carbonilação Proteica/fisiologia , Diálise Renal , Idoso , Nefropatias Diabéticas/sangue , Feminino , Humanos , Falência Renal Crônica/sangue , Masculino , Estresse Oxidativo/fisiologia
10.
Parasitol Res ; 117(9): 2957-2962, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29987413

RESUMO

Acanthamoeba has 22 genotypes with the T4 genotype being the main causative agent of amoebic granulomatous encephalitis and keratitis. Because the molecular mechanisms of the immune defenses of neutrophils and macrophages against histoparasites are based on oxidative stress, parasites may rely on their antioxidant systems to preclude immune defenses. Therefore, understanding of the effect of oxidative stress on vital characteristics of Acanthamoeba castellanii (T4 genotype) and the antioxidant defense responses of Acanthamoeba to oxidative status will cast light on immune cell-parasite interactions. Acanthamoeba T4 cells were cultured in RPMI-1640 medium containing different concentrations of hydrogen peroxide (H2O2). The survival of Acanthamoeba was evaluated by MTT assay and the IC50 concentration was calculated. The total antioxidant capacity (TAC) of the parasite was determined by the cupric reducing antioxidant capacity (CUPRAC) method. Malondialdehyde (MDA) as a marker of lipid peroxidation, protein carbonyl content as a measure of oxidized protein, total thiol (-SH) groups present on proteins as a major source of cellular antioxidants, and total oxidant status (TOS) were evaluated by colorimetric methods. The reactive oxygen species level increased markedly after induction of oxidative stress by the treatment of Acanthamoeba T4 with H2O2. Exposure to H2O2 also significantly increased the MDA and protein carbonyl content. The TOS level and total thiol groups also increased in the treated group compared to those in untreated parasites, although the results were not statistically significant. The TAC level was found to be significantly higher in H2O2-treated parasites, confirming that the parasite fosters its total antioxidant capacity to overcome oxidative conditions. This study showed that under oxidative stress, the defense reactions of the parasite are in part mediated by increasing its antioxidant activity, which is important for the survival of the parasite.


Assuntos
Acanthamoeba castellanii/metabolismo , Antioxidantes/metabolismo , Peróxido de Hidrogênio/farmacologia , Estresse Oxidativo/fisiologia , Acanthamoeba castellanii/genética , Biomarcadores , Genótipo , Humanos , Peroxidação de Lipídeos/fisiologia , Malondialdeído/análise , Oxirredução/efeitos dos fármacos , Carbonilação Proteica/fisiologia , Proteólise
11.
J Pharm Sci ; 107(10): 2570-2580, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29935298

RESUMO

Protein carbonylation is a posttranslational modification referring to the occurrence of aldehydes and ketones in proteins. The current understanding of how carbonylation, in particular, metal-catalyzed carbonylation, occurs in recombinant mAbs during production and storage is very limited. To facilitate investigations into mAb carbonylation, we developed a protein carbonylation assay with improved assay robustness and precision over the conventional assays. We applied this assay to investigate mAb carbonylation under production, storage, and stress conditions and showed that iron, hydrogen peroxide, and polysorbate 20 at pharmaceutically relevant levels critically influence the extent of mAb carbonylation. In addition, we found that while carbonylation correlates with mAb aggregation in several cases, carbonylation cannot be used as a general indicator for aggregation. Furthermore, we observed that mAb carbonylation level can decrease during storage, which indicates that carbonylation products may not be stable. Finally, we report for the first time a positive correlation between carbonylation and acidic charge heterogeneity of mAbs that underwent metal-catalyzed oxidation. This finding shows that the impact of protein carbonylation on product quality for mAbs is not limited to aggregation but also extends to charge heterogeneity.


Assuntos
Anticorpos Catalíticos/química , Anticorpos Monoclonais/química , Metais/química , Proteínas/química , Bioensaio/métodos , Catálise , Peróxido de Hidrogênio/química , Oxirredução , Carbonilação Proteica/fisiologia
12.
Alcohol Clin Exp Res ; 42(7): 1192-1205, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29708596

RESUMO

BACKGROUND: Glutathione S-transferase A4-4 (GSTA4) is a key enzyme for removal of toxic lipid peroxidation products such as 4-hydroxynonenal (4-HNE). In this study, we examined the potential role of GSTA4 on protein carbonylation and progression of alcoholic liver disease by examining the development of liver injury in male wild-type (WT) SV/J mice and SV/J mice lacking functional GSTA4 (GSTA4-/- mice). METHODS: Adult male WT and GSTA4-/- mice were fed chow (N = 10 to 12) or high-fat Lieber-DeCarli liquid diets containing up to 28% calories as ethanol (EtOH) (N = 18 to 20) for 116 days. At the end of the study, half of the EtOH-fed mice were acutely challenged with an EtOH binge (3 g/kg given intragastrically) 12 hours before sacrifice. Carbonylation of liver proteins was assessed by immunohistochemical staining for 4-HNE adduction and by comprehensive liquid chromatography-tandem mass spectrometry (LC-MS/MS) of purified carbonylated proteins. RESULTS: Chronic EtOH intake significantly increased hepatic 4-HNE adduction and protein carbonylation, including carbonylation of ribosomal proteins. EtOH intake also resulted in steatosis and increased serum alanine aminotransferase. Hepatic infiltration with B cells, T cells, and neutrophils and mRNA expression of pro-inflammatory cytokines tumor necrosis factor (TNF)α and interferon (IFN)γ was modest in WT mice. However, an EtOH binge increased hepatic necrosis, hepatic cell proliferation, and expression of TNFα mRNA (p < 0.05). EtOH treatment of GSTA4-/- mice increased B-cell infiltration and increased mRNA expression of TNFα and IFNγ and of matrix remodeling markers MMP9, MMP13, and Col1A1 (p < 0.05). GSTA4-/- mice exhibited panlobular rather than periportal distribution of 4-HNE-adducted proteins and increased overall 4-HNE staining after EtOH binge. Comprehensive LC-MS of carbonylated proteins identified 1,022 proteins of which 189 were unique to the GSTA4-/- group. CONCLUSIONS: These data suggest long-term adaptation to EtOH in WT mice does not occur in GSTA4-/- mice. Products of lipid peroxidation appear to play a role in inflammatory responses due to EtOH. And EtOH effects on B-cell infiltration and autoimmune responses may be secondary to formation of carbonyl adducts.


Assuntos
Etanol/toxicidade , Glutationa Transferase/deficiência , Glutationa Transferase/genética , Hepatopatias Alcoólicas/genética , Hepatopatias Alcoólicas/metabolismo , Carbonilação Proteica/fisiologia , Animais , Etanol/administração & dosagem , Glutationa Transferase/química , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Hepatopatias Alcoólicas/patologia , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Knockout , Carbonilação Proteica/efeitos dos fármacos , Estrutura Secundária de Proteína
13.
Aging (Albany NY) ; 10(5): 868-901, 2018 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-29779015

RESUMO

Non-enzymatic protein modifications occur inevitably in all living systems. Products of such modifications accumulate during aging of cells and organisms and may contribute to their age-related functional deterioration. This review presents the formation of irreversible protein modifications such as carbonylation, nitration and chlorination, modifications by 4-hydroxynonenal, removal of modified proteins and accumulation of these protein modifications during aging of humans and model organisms, and their enhanced accumulation in age-related brain diseases.


Assuntos
Envelhecimento/metabolismo , Encefalopatias/metabolismo , Encefalopatias/fisiopatologia , Estresse Oxidativo/fisiologia , Processamento de Proteína Pós-Traducional/fisiologia , Animais , Halogenação/fisiologia , Humanos , Carbonilação Proteica/fisiologia
14.
J Psychiatr Res ; 102: 29-33, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29574402

RESUMO

BACKGROUND: Oxidative stress has been implicated in the pathophysiology of mood disorders in young adults. However, there is few data to support its role in the elderly. The primary aim of this study was to evaluate whether subjects with late-life depression (LLD) presented with changes in oxidative stress response in comparison with the non-depressed control group. We then explored how oxidative stress markers associated with specific features of LLD, in particular cognitive performance and age of onset of major depressive disorder in these individuals. METHODS: We included a convenience sample of 124 individuals, 77 with LLD and 47 non-depressed subjects (Controls). We measure the plasma levels of 6 oxidative stress markers: thiobarbituric acid reactive substances (TBARS), protein carbonil content (PCC), free 8-isoprostane, glutathione peroxidase (GPx) activity, glutathione reductase (GR) activity, and glutathione S-transferase (GST) activity. RESULTS: We found that participants with LLD had significantly higher free 8-isoprostane levels (p = 0.003) and lower glutathione peroxidase activity (p = 0.006) compared to controls. Free 8-isoprostane levels were also significantly correlated with worse scores in the initiation/perseverance (r = -0.24, p = 0.01), conceptualization (r = -0.22, p = 0.02) sub-scores, and the total scores (r = -0.21, p = 0.04) on the DRS. CONCLUSIONS: Our study provides robust evidence of the imbalance between oxidative stress damage, in particular lipid peroxidation, and anti-oxidative defenses as a mechanism related to LLD, and cognitive impairment in this population. Interventions aiming to reduce oxidative stress damage can have a potential neuroprotective effect for LLD subjects.


Assuntos
Depressão/metabolismo , Depressão/fisiopatologia , Peroxidação de Lipídeos/fisiologia , Estresse Oxidativo/fisiologia , Idoso , Idoso de 80 Anos ou mais , Transtornos Cognitivos/etiologia , Depressão/complicações , F2-Isoprostanos/metabolismo , Feminino , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Glutationa Transferase/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Carbonilação Proteica/fisiologia , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo
15.
J Neurol Sci ; 387: 85-91, 2018 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-29571878

RESUMO

Misfolded protein aggregates are the hallmark of Amyotrophic Lateral Sclerosis (ALS) which suggests involvement of protein homeostasis pathways in etiology of ALS. However, status of protein homeostasis in peripheral blood of ALS is not well established. We analyzed expression levels of key genes of proteostasis pathways in peripheral blood mononuclear cells (PBMCs) of sporadic ALS (sALS) patients and healthy controls. Increased protein carbonylation was observed in patients reflecting oxidative damage in PBMCs. We observed increased transcript and protein levels of GRP78 suggesting Endoplasmic reticulum (ER) insult to cells. Further, significant upregulation of spliced XBP1 and two stress sensors: IRE1α/ERN1 and ATF6 indicated induction of unfolded protein response (UPR). Genes involved in autophagosome initiation (ULK1, ULK2, ATG13); nucleation and elongation (BECLIN1, ATG7, ATG16L1, ATG5, ATG10) and vesicular trafficking genes were significantly increased in patients. Increased lipidation of LC3 validated induction of autophagy. Accumulation of low molecular weight ubiquitinated proteins in patients suggested deregulation of proteasome (UPS) pathway. In addition, cytosolic chaperones (HSP70 and HSP27) and HSF1 were elevated in patients. Increased TDP43 indicated role of TDP43 in disease pathology. Our findings suggest that there is oxidative insult and upregulation of UPR, vesicular trafficking and autophagy in PBMCs of sALS patients.


Assuntos
Esclerose Amiotrófica Lateral/patologia , Esclerose Amiotrófica Lateral/fisiopatologia , Leucócitos Mononucleares/metabolismo , Proteostase/fisiologia , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologia , Adulto , Idoso , Autofagia/fisiologia , Estresse do Retículo Endoplasmático/fisiologia , Feminino , Expressão Gênica/fisiologia , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Carbonilação Proteica/fisiologia , Desdobramento de Proteína , Proteínas Ubiquitinadas/genética , Proteínas Ubiquitinadas/metabolismo , Proteína 1 de Ligação a X-Box/genética , Proteína 1 de Ligação a X-Box/metabolismo
16.
J Neuroinflammation ; 15(1): 71, 2018 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-29514678

RESUMO

BACKGROUND: In multiple sclerosis (MS), neurodegeneration is the main reason for chronic disability. Alpha-lipoic acid (LA) is a naturally occurring antioxidant which has recently been demonstrated to reduce the rate of brain atrophy in progressive MS. However, it remains uncertain if it is also beneficial in the early, more inflammatory-driven phases. As clinical studies are costly and time consuming, optic neuritis (ON) is often used for investigating neuroprotective or regenerative therapeutics. We aimed to investigate the prospect for success of a clinical ON trial using an experimental autoimmune encephalomyelitis-optic neuritis (EAE-ON) model with visual system readouts adaptable to a clinical ON trial. METHODS: Using an in vitro cell culture model for endogenous oxidative stress, we compared the neuroprotective capacity of racemic LA with the R/S-enantiomers and its reduced form. In vivo, we analyzed retinal neurodegeneration using optical coherence tomography (OCT) and the visual function by optokinetic response (OKR) in MOG35-55-induced EAE-ON in C57BL/6J mice. Ganglion cell counts, inflammation, and demyelination were assessed by immunohistological staining of retinae and optic nerves. RESULTS: All forms of LA provided equal neuroprotective capacities in vitro. In EAE-ON, prophylactic LA therapy attenuated the clinical EAE score and prevented the thinning of the inner retinal layer while therapeutic treatment was not protective on visual outcomes. CONCLUSIONS: A prophylactic LA treatment is necessary to protect from visual loss and retinal thinning in EAE-ON, suggesting that a clinical ON trial starting therapy after the onset of symptoms may not be successful.


Assuntos
Encefalomielite Autoimune Experimental/patologia , Degeneração Neural/prevenção & controle , Retina/patologia , Ácido Tióctico/uso terapêutico , Transtornos da Visão/prevenção & controle , Complexo Vitamínico B/uso terapêutico , Animais , Complexo CD3/metabolismo , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/induzido quimicamente , Encefalomielite Autoimune Experimental/complicações , Feminino , Glutationa/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteína Básica da Mielina/metabolismo , Degeneração Neural/etiologia , Nistagmo Optocinético/fisiologia , Carbonilação Proteica/fisiologia , Tomografia de Coerência Óptica , Transtornos da Visão/etiologia
17.
Neurogastroenterol Motil ; 30(7): e13305, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29411462

RESUMO

BACKGROUND: Anorexia nervosa, a restrictive eating disorder, is often associated with gastrointestinal disorders, particularly a delayed gastric emptying. However, the mechanisms remained poorly documented. Thus, we aimed to evaluate gastric emptying and antrum protein metabolism in the Activity-Based Anorexia model (ABA). METHODS: Females C57Bl/6 mice were randomized into 3 groups: Control, ABA, and Limited Food Access (LFA). Food access has been progressively limited from 6 h/day at day 6 to 3 h/day at day 9 and until day 17. ABA mice had free access to an activity wheel. Gastric emptying was assessed. On gastric extracts, a proteomic analysis was performed, as well as an evaluation of protein synthesis and protein oxidation. KEY RESULTS: Both LFA and ABA mice exhibited a delayed gastric emptying compared with Controls (P < .05). Proteomic approach revealed 15 proteins that were differentially expressed. Among these proteins, we identified 2 clusters of interest contributing to (i) the organization of muscle fiber with ACTA2, VCL, KRT19, KRT8, and DES proteins and (ii) "heat shock proteins" with STIP1, HSPD1, and HSPA8 proteins. ABA mice specifically exhibited an increased rate of gastric oxidized proteins. CONCLUSIONS AND INFERENCES: Delayed gastric emptying observed in anorectic conditions appears to be secondary to malnutrition. However, an oxidative stress is specifically present in the stomach of ABA mice. Its role remains to be further studied.


Assuntos
Anorexia/metabolismo , Esvaziamento Gástrico/fisiologia , Gastroparesia/metabolismo , Carbonilação Proteica/fisiologia , Antro Pilórico/metabolismo , Animais , Anorexia/complicações , Anorexia/fisiopatologia , Feminino , Gastroparesia/etiologia , Gastroparesia/fisiopatologia , Camundongos , Camundongos Endogâmicos C57BL , Distribuição Aleatória , Espectrometria de Massas por Ionização por Electrospray/métodos
18.
FEBS Lett ; 592(6): 1010-1019, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29430658

RESUMO

Reactive carbonyls (RCs), which are inevitably produced during respiratory and photosynthetic metabolism, have the potential to cause oxidative damage to photosynthetic organisms. Previously, we proposed a scavenging model for RCs in the cyanobacterium Synechocystis sp. PCC 6803 (S. 6803). In the current study, we constructed mutants deficient in the enzymes medium-chain dehydrogenase/reductase (ΔMDR) and aldo-keto reductase (ΔAKR) to investigate their contributions to RC scavenging in vivo. We found that treatment with the lipid-derived RC acrolein causes growth inhibition and promotes greater protein carbonylation in ΔMDR, compared with the wild-type and ΔAKR. In both ΔMDR and ΔAKR, photosynthesis is severely inhibited in the presence of acrolein. These results suggest that these enzymes function as part of the scavenging systems for RCs in S. 6803 in vivo.


Assuntos
Acil-CoA Desidrogenase/metabolismo , Aldo-Ceto Redutases/metabolismo , Proteínas de Bactérias/metabolismo , Radicais Livres/metabolismo , Synechocystis/enzimologia , Acil-CoA Desidrogenase/genética , Aldo-Ceto Redutases/genética , Proteínas de Bactérias/genética , Fotossíntese/fisiologia , Carbonilação Proteica/fisiologia , Synechocystis/genética
19.
Biomed Res Int ; 2018: 2828143, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30662904

RESUMO

To research carbonylated proteins and screen molecular targets in the rat striatum on regular aerobic exercise, male Sprague-Dawley rats (13 months old, n = 24) were randomly divided into middle-aged sedentary control (M-SED) and aerobic exercise (M-EX) groups (n = 12 each). Maximum oxygen consumption (VO2max) gradually increased from 50%-55% to 65%-70% for a total of 10 weeks. A total of 36 carbonylated proteins with modified oxidative sites were identified by Electrospray Ionization-Quadrupole-Time of Flight-Mass Spectrometer (ESI-Q-TOF-MS), including 17 carbonylated proteins unique to the M-SED group, calcium/calmodulin-dependent protein kinase type II subunit beta (CaMKIIß), and heterogeneous nuclear ribonucleoprotein A2/B1 (Hnrnpa2b1), among others, and 19 specific to the M-EX group, ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1), and malic enzyme, among others. Regular aerobic exercise improved behavioral and stereological indicators, promoted normal apoptosis (P < 0.01), alleviated carbonylation of the CaMKIIß and Hnrnpa2b1, but induced carbonylation of the UCH-L1, and significantly upregulated the expression levels of CaMKIIß, CaMKIIα, and Vdac1 (p < 0.01) and Hnrnpa2b1 and UCH-L1 (p < 0.01), as well as the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathways (PI3K/Akt/mTOR) pathway-related genes Akt and mTOR. Regular aerobic exercise for 10 weeks (incremental for the first 6 weeks followed by constant loading for 4 weeks) enhanced carbonylation of CaMKIIß, Hnrnpa2b1, and modulated apoptosis via activation of CaMK and phosphoinositide 3-kinase/protein kinase B/mTOR signaling. It also promoted normal apoptosis in the rat striatum, which may have protective effects in neurons.


Assuntos
Apoptose/fisiologia , Proteínas de Transporte/metabolismo , Condicionamento Físico Animal/fisiologia , Carbonilação Proteica/fisiologia , Transdução de Sinais/fisiologia , Animais , Proteínas de Ligação ao Cálcio , Masculino , Oxirredução , Estresse Oxidativo/fisiologia , Consumo de Oxigênio/fisiologia , Fosfatidilinositol 3-Quinase/metabolismo , Proteômica/métodos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Ratos Sprague-Dawley , Serina-Treonina Quinases TOR/metabolismo , Ubiquitina Tiolesterase/metabolismo , Regulação para Cima/fisiologia
20.
Neurochem Res ; 43(3): 609-618, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29264677

RESUMO

This study was conducted to further our understanding about the link between lipid peroxidation and protein carbonylation in rat brain slices incubated with the glutathione (GSH)-depletor diethyl maleate. Using this in vitro system of oxidative stress, we found that there is a significant lag between the appearance of carbonylated proteins and GSH depletion, which seems to be due to the removal of oxidized species early on in the incubation by the mitochondrial Lon protease. Upon acute GSH depletion, protein carbonyls accumulated mostly in mitochondria and to a lesser degree in other subcellular fractions that also contain high levels of polyunsaturated lipids. This result is consistent with our previous findings suggesting that lipid hydroperoxides mediate the oxidation of proteins in this system. However, these lipid hydroperoxides are not produced by oxidation of free arachidonic acid or other polyunsaturated free fatty acids by lipooxygenases or cyclooxygenases. Finally, γ-glutamyl semialdehyde and 2-amino-adipic semialdehyde were identified by HPLC as the carbonyl-containing amino acid residues, indicating that proteins are carbonylated by metal ion-catalyzed oxidation of lysine, arginine and proline residues. The present findings are important in the context of neurological disorders that exhibit increased lipid peroxidation and protein carbonylation, such as Parkinson's disease, Alzheimer's disease, and multiple sclerosis.


Assuntos
Encéfalo/metabolismo , Glutationa/deficiência , Peroxidação de Lipídeos/fisiologia , Carbonilação Proteica/fisiologia , Animais , Glutationa/metabolismo , Peróxidos Lipídicos/metabolismo , Masculino , Mitocôndrias/metabolismo , Estresse Oxidativo/fisiologia , Ratos Sprague-Dawley , Frações Subcelulares/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA