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1.
Clin Exp Nephrol ; 28(7): 674-682, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38457030

RESUMO

BACKGROUND: Dialysis patients are susceptible to developing severe coronavirus disease 2019 (COVID-19) due to hypoimmunity. Antibody titers against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) after the primary vaccinations are lower in hemodialysis (HD) patients than in healthy individuals. This study aimed to evaluate the effect of a SARS-CoV-2 booster vaccination in HD and peritoneal dialysis (PD) patients based on antibody titers and cellular and humoral immunity. METHODS: Participants of the control, HD, and PD groups were recruited from 12 facilities. SARS-CoV-2 antigen-specific cytokine and IgG-antibody levels were measured. Regulatory T cells and memory B cells were counted using flow cytometry at 6 months after primary vaccination with BNT162b2 and 3 weeks after the booster vaccination in HD and PD patients and compared with those of a control group. RESULTS: Booster vaccination significantly enhanced the levels of antibodies, cytokines, and memory B cells in three groups. The HD group showed significantly higher levels of IgG-antibodies, IL-1ß, IL-2, IL-4, IL-17, and memory B cells than those in the control group at 3 weeks after the booster dose. The PD group tended to show similar trends to HD patients but had similar levels of IgG-antibodies, cytokines, and memory B cells to the control group. CONCLUSIONS: HD patients had significantly stronger cellular and humoral immune responses than the control 3 weeks after the booster dose. Our findings will help in developing better COVID-19 vaccination strategies for HD and PD patients.


Assuntos
Anticorpos Antivirais , Vacina BNT162 , COVID-19 , Imunidade Humoral , Imunização Secundária , Diálise Renal , Humanos , Masculino , Feminino , COVID-19/imunologia , COVID-19/prevenção & controle , Pessoa de Meia-Idade , Idoso , Anticorpos Antivirais/sangue , Vacina BNT162/imunologia , Citocinas/sangue , SARS-CoV-2/imunologia , Vacinas contra COVID-19/imunologia , Imunidade Celular , Imunoglobulina G/sangue , Japão , Células B de Memória/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Diálise Peritoneal , População do Leste Asiático
2.
ERJ Open Res ; 9(2)2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37009022

RESUMO

Cough is a major symptom in patients with asthma and poses a significant burden compared with other asthma symptoms. However, there are no approved treatments in Japan, developed to specifically treat cough in patients with asthma. We present the design of REACH, an 8-week real-life study, which will evaluate the efficacy of a combination of indacaterol acetate, glycopyrronium bromide and mometasone furoate (IND/GLY/MF) in asthmatic patients with cough refractory to medium-dose inhaled corticosteroid/long-acting ß2-agonist (ICS/LABA). Patients with asthma (age ≥20 to <80 years) with a cough visual analogue scale (VAS) ≥40 mm will be randomised 2:1:1 to receive IND/GLY/MF medium-dose 150/50/80 µg once daily or step-up to a high-dose regimen of fluticasone furoate/vilanterol trifenatate (FF/VI) 200/25 µg once daily or budesonide/formoterol fumarate (BUD/FM) 160/4.5 µg four inhalations twice daily during the 8-week treatment period. The primary objective is to demonstrate the superiority of IND/GLY/MF medium-dose over high-dose ICS/LABA in terms of cough-specific quality of life after 8 weeks. The key secondary objective is to demonstrate the superiority of IND/GLY/MF in terms of subjective assessment of cough severity. Cough frequency (VitaloJAK cough monitor) and capsaicin cough receptor sensitivity will be evaluated in eligible patients. Cough VAS scores, fractional exhaled nitric oxide, spirometry and blood tests, and the Asthma Control Questionnaire-6, Cough and Sputum Assessment Questionnaire, and Japanese version of the Leicester Cough Questionnaire will be evaluated. REACH will provide valuable evidence on whether a switch to IND/GLY/MF medium-dose or step-up to high-dose ICS/LABA is beneficial for patients with persistent cough despite treatment with medium-dose ICS/LABA.

3.
Antioxid Redox Signal ; 9(9): 1427-37, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17627468

RESUMO

Human thioredoxin-1 (hTrx) exhibits a disulfide reducing activity and was originally identified as a soluble cytokine-like factor secreted from cells of a human T-cell leukemia virus type I (HTLV-I)-transformed cell line. Recent studies have revealed that endogenous Trx plays an important role in cytoprotection against various oxidative stress-associated disorders. However, the function of exogenous Trx is still not fully understood. We report here that a cysteine-modified mutant of recombinant human Trx (rhTrx-C35S) binds to human umbilical vein endothelial cells (HUVECs) as well as stimulated T cells and rapidly enters these cells via lipid rafts. In addition, we found that endogenous Trx is expressed on the surface of HUVECs, including lipid rafts. These events suggest cell-surface Trx as a possible target of rhTrx-C35S. Furthermore, we found that anti-human Trx mouse monoclonal antibody inhibits adherence of LPS-stimulated human peripheral blood polymorphonuclear cells (PMNs) to HUVECs. This adherence was also suppressed by a recombinant human Trx (rhTrx), but not by a mutant rhTrx (rhTrx-C32S/C35S) with no reducing activity. Cell-surface Trx may be involved in the process of interaction between PMNs and HUVECs and a possible target of cysteine-modified exogenous Trx as well as wild-type exogenous Trx through redox regulation.


Assuntos
Endotélio Vascular/fisiologia , Leucócitos/fisiologia , Microdomínios da Membrana/fisiologia , Proteínas de Membrana/fisiologia , Tiorredoxinas/metabolismo , Linhagem Celular Transformada , Endotélio Vascular/efeitos dos fármacos , Vírus Linfotrópico T Tipo 1 Humano/genética , Humanos , Células Jurkat , Leucócitos/efeitos dos fármacos , Microdomínios da Membrana/ultraestrutura , Microscopia Confocal , Proteínas Recombinantes/farmacologia , Tiorredoxinas/genética
4.
Antioxid Redox Signal ; 9(9): 1439-48, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17627472

RESUMO

Thioredoxin-1 (TRX) plays important roles in cellular signaling by controlling the redox state of cysteine residues in target proteins. TRX is released in response to oxidative stress and shows various biologic functions from the extracellular environment. However, the mechanism by which extracellular TRX transduces the signal into the cells remains unclear. Here we report that the cysteine modification at the active site of TRX promotes the internalization of TRX into the cells. TRX-C35S, in which the cysteine at residue 35 of the active site was replaced with serine, was internalized more effectively than wild-type TRX in human T-cell leukemia virus-transformed T cells. TRX-C35S bound rapidly to the cell surface and was internalized into the cells dependent on lipid rafts in the plasma membrane. This process was inhibited by wild-type TRX, reducing reagents such as dithiothreitol, and methyl-beta-cyclodextrin, which disrupts lipid rafts. Moreover, the internalized TRX-C35S binds to endogenous TRX, resulting in the generation of intracellular reactive oxygen species (ROS) and enhanced cis-diamine-dichloroplatinum (II) (CDDP)-induced apoptosis via a ROS-mediated pathway involving apoptosis signal-regulating kinase-1 (ASK-1) activation. These findings suggest that the cysteine at the active site of TRX plays a key role in the internalization and signal transduction of extracellular TRX into the cells.


Assuntos
Microdomínios da Membrana/fisiologia , Tiorredoxinas/metabolismo , Substituição de Aminoácidos , Apoptose , Cisteína , Humanos , Células Jurkat , Modelos Biológicos , Proteínas Recombinantes/metabolismo , Tiorredoxinas/antagonistas & inibidores , Tiorredoxinas/genética
5.
Free Radic Res ; 41(8): 861-9, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17654042

RESUMO

Indomethacin is one of non-steroidal anti-inflammatory drugs that are commonly used clinically and often cause gastric mucosal injury as a side effect. Generation of reactive oxygen species (ROS) and activation of apoptotic signaling are involved in the pathogenesis of indomethacin-induced gastric mucosal injury. Thioredoxin-1 (Trx-1) is a small redox-active protein with anti-oxidative activity and redox-regulating functions. The aim of this study was to investigate the protective effect of Trx-1 against indomethacin-induced gastric mucosal injury. Trx-1 transgenic mice displayed less gastric mucosal damage than wild type (WT) C57BL/6 mice after intraperitoneal administration of indomethacin. Administration of recombinant human Trx-1 (rhTrx-1) or transfection of the Trx-1 gene reduced indomethacin-induced cytotoxicity in rat gastric epithelial RGM-1 cells. Pretreatment with rhTrx-1 suppressed indomethacininduced ROS production and downregulation of phosphorylated Akt in RGM-1 cells. Survivin, a member of inhibitors of apoptosis proteins family, was downregulated by indomethacin, which was suppressed in Trx-1 transgenic mice or by administration of rhTrx-1 in RGM-1 cells. Trx-1 inhibits indomethacin-induced apoptotic signaling and gastric ulcer formation, suggesting that it may have a preventive and therapeutic potential against indomethacin-induced gastric injury.


Assuntos
Anti-Inflamatórios não Esteroides/antagonistas & inibidores , Mucosa Gástrica/efeitos dos fármacos , Indometacina/antagonistas & inibidores , Tiorredoxinas/farmacologia , Animais , Anti-Inflamatórios não Esteroides/toxicidade , Apoptose/efeitos dos fármacos , Apoptose/genética , Linhagem Celular , Mucosa Gástrica/metabolismo , Indometacina/toxicidade , Camundongos , Camundongos Transgênicos , Ratos , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Tiorredoxinas/genética , Tiorredoxinas/metabolismo
6.
J Neurosci ; 25(9): 2396-404, 2005 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-15745966

RESUMO

Exposure to excessive light induces retinal photoreceptor cell damage, leading to development and progression of various retinal diseases. We tested the effect of geranylgeranylacetone (GGA), an acyclic polyisoprenoid, on light-induced retinal damage in mice. Oral treatment with GGA (1.0 mg/d) for 5 d induced thioredoxin (Trx) and heat shock protein 72 (Hsp72) predominantly in the retinal pigment epithelium (RPE). After white light exposure (8000 lux for 2 h), the percentage of terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive photoreceptor cells decreased significantly at 24 and 96 h, and the number of photoreceptor cell nuclei at 96 h and the electroretinographic amplitudes of the a- and b-waves at 4 and 10 d increased significantly in GGA-pretreated mice compared with saline-pretreated mice. Light-induced upregulations of 8-hydroxy-2-deoxyguanosine and 4-hydroxy-2-nonenal-modified protein, markers of oxidative stress, were inhibited by GGA pretreatment. To elucidate the cytoprotective mechanism of GGA and Trx, we used human K-1034 RPE cells and mouse photoreceptor-derived 661W cells. In K-1034 cells, GGA (10 microM) induced intracellular Trx, Hsp72, and extracellular Trx but not extracellular Hsp72. Extracellular Trx (0.75 nM) attenuated H2O2 (200 microM)-induced cell damage in 661W cells. Pretreatment with GGA and overexpression of Trx in K-1034 cells counteracted H2O2 (50 microM)-induced attenuation of cellular latex bead incorporation. Protection of phagocytotic activity through induction of Trx and possibly Hsp72 in RPE cells and elimination of oxidative stress in the photoreceptor layer through release of Trx from RPE cells may be mechanisms of GGA-mediated cytoprotection. Therefore, Trx is a neurotrophic factor released from RPE cells and plays a crucial role in maintaining photoreceptor cell integrity.


Assuntos
Diterpenos/uso terapêutico , Fármacos Neuroprotetores/uso terapêutico , Células Fotorreceptoras/efeitos dos fármacos , Doenças Retinianas/tratamento farmacológico , Doenças Retinianas/patologia , 8-Hidroxi-2'-Desoxiguanosina , Aldeídos/metabolismo , Animais , Western Blotting/métodos , Contagem de Células , Morte Celular/efeitos dos fármacos , Linhagem Celular , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Modelos Animais de Doenças , Eletrorretinografia/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Citometria de Fluxo/métodos , Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP72/metabolismo , Humanos , Peróxido de Hidrogênio/farmacologia , Imuno-Histoquímica/métodos , Marcação In Situ das Extremidades Cortadas/métodos , Luz/efeitos adversos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Neoplasias/metabolismo , Fagócitos/efeitos dos fármacos , Células Fotorreceptoras/efeitos da radiação , Doenças Retinianas/fisiopatologia , Tiorredoxinas/metabolismo , Fatores de Tempo
7.
Antioxid Redox Signal ; 8(9-10): 1881-90, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16987040

RESUMO

Oxidative stresses are largely mediated by intracellular protein oxidations by reactive oxygen species (ROS). Host cells are equipped with antioxidants that scavenge ROS. The cellular reduction/oxidation (redox) balance is maintained by ROS and antioxidants. Accumulating evidence suggests that the redox balance plays an important role in cellular signaling through the redox modification of cysteine residues in various important components of the signal transduction pathway. Thioredoxin (TRX) is a small protein playing important roles in cellular responses, including cell growth, cell cycle, gene expression, and apoptosis, to maintain the redox circumstance. Moreover, many recent papers have shown that the redox regulation by TRX is deeply involved in the pathogenesis of various oxidative stress-associated disorders. This review focuses on TRX and its related molecules, and discusses the role of TRX-dependent redox regulation in oxidative stress-induced signal transduction.


Assuntos
Transdução de Sinais/fisiologia , Tiorredoxinas/metabolismo , Humanos , Modelos Biológicos , Oxirredução , Estresse Oxidativo/fisiologia , Processamento de Proteína Pós-Traducional/fisiologia , Espécies Reativas de Oxigênio/metabolismo
8.
Antioxid Redox Signal ; 8(9-10): 1891-6, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16987041

RESUMO

Thioredoxin-1 (TRX) is a small redox-active protein with antioxidative effects and redox-regulating functions. Cigarette smoking is a major etiological factor in the pathogenesis of a variety of diseases and recruits systemic immune and inflammatory responses. This report demonstrates that TRX attenuates the systemic inflammatory responses induced by cigarette smoking. The mRNA expressions of tumor necrosis factor alpha (TNF-alpha) and macrophage migration inhibitory factor (MIF) were suppressed in the spleen of TRX overexpressing transgenic mice (TRX-tg) exposed to cigarette smoking, compared with control C57BL/6 mice. In addition, protein carbonylation, a marker of cellular protein oxidation, was enhanced by cigarette smoking in the tissues of heart and liver in control mice more than in TRX-tg mice. These findings suggest that TRX may suppress the systemic inflammatory responses against cigarette smoking.


Assuntos
Inflamação/metabolismo , Fumar/metabolismo , Tiorredoxinas/genética , Animais , Humanos , Fígado/metabolismo , Fatores Inibidores da Migração de Macrófagos/genética , Fatores Inibidores da Migração de Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miocárdio/metabolismo , Oxirredução , Estresse Oxidativo/fisiologia , Carbonilação Proteica/fisiologia , Proteínas/metabolismo , Baço/metabolismo , Tiorredoxinas/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
9.
Cell Res ; 16(2): 230-9, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16474438

RESUMO

Thioredoxin-1 (TRX) is a stress-inducible redox-regulatory protein with antioxidative and anti-inflammatory effects. Here we show that the release of histamine from mast cells elicited by cross-linking of high-affinity receptor for IgE (FcepsilonRI) was significantly suppressed in TRX transgenic (TRX-tg) mice compared to wild type (WT) mice. Intracellular reactive oxygen species (ROS) of mast cells stimulated by IgE and antigen was also reduced in TRX-tg mice compared to WT mice. Whereas there was no difference in the production of cytokines (IL-6 and TNF-alpha) from mast cells in response to 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) stimulation in TRX-tg and WT mice. Immunological status of TRX-tg mice inclined to T helper (Th) 2 dominant in primary immune response, although there was no difference in the population of dendritic cells (DCs) and regulatory T cells. We conclude that the histamine release from mast cells in TRX-tg mice is suppressed by inhibition of ROS generation. As ROS are involved in mast cell activation and facilitate mediator release, TRX may be a key signaling molecule regulating the early events in the IgE signaling in mast cells and the allergic inflammation.


Assuntos
Liberação de Histamina , Mastócitos/metabolismo , Isoformas de Proteínas/metabolismo , Tiorredoxinas/metabolismo , Animais , Bovinos , Células Dendríticas/metabolismo , Dinitrofenóis/imunologia , Haptenos/imunologia , Humanos , Interleucina-6/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Oxirredução , Isoformas de Proteínas/genética , Espécies Reativas de Oxigênio/metabolismo , Receptores de IgE/metabolismo , Soroalbumina Bovina/imunologia , Baço/citologia , Tiorredoxinas/genética , Fator de Necrose Tumoral alfa/metabolismo
10.
Biofactors ; 27(1-4): 47-51, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17012763

RESUMO

Thioredoxin (TRX) binding protein-2 (TBP-2), a negative regulator of TRX, is involved in intracellular redox regulation and cellular growth. The expression of TBP-2 is frequently lost in tumor cell lines and tissues, whereas the ectopic expression of TBP-2 suppresses cellular proliferation along with cell cycle arrest at the G1 phase. TBP-2 was also reported to be a cellular senescence-associated gene. Besides the retardation of cellular growth, the reduction of white adipose, and alteration of the energy pathway are involved in several features of the aging process. We have generated TBP-2 genetically modified mice and found that TBP-2 is closely linked to lipid metabolism. Indeed, TBP-2 has been suggesting to be related to familial combined hyperlipidemia analyzed by a spontaneous mutant mouse strain. As lipid metabolism is one of the most primitive sources of energy production, we discussed the possible roles of TBP-2 in the regulation of energy utilization connected to the aging process.


Assuntos
Proteínas de Transporte/fisiologia , Senescência Celular/fisiologia , Animais , Apoptose/fisiologia , Ciclo Celular/fisiologia , Metabolismo Energético/fisiologia , Humanos , Modelos Biológicos , Oxirredução , Tiorredoxinas
11.
Oncogene ; 22(12): 1860-5, 2003 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-12660821

RESUMO

Thioredoxin is a redox-regulating protein, the expression of which is induced by various forms of oxidative stress. Thioredoxin controls the interactions of various transcription factors through redox regulation. In K562 cells, we have previously reported that hemin induces activation of the thioredoxin gene by regulating NF-E2-related factor (Nrf2) through the antioxidant responsive element (ARE). We showed here that tert-butylhydroquinone (tBHQ), an electrophile stressor, activates the thioredoxin gene through the ARE. In an electrophoretic mobility shift assay, a specific Nrf2/small Maf binding complex was induced by tBHQ and bound to the ARE. Overexpression of Nrf2 increased the tBHQ-induced thioredoxin gene activation through the ARE, whereas that of Jun and Fos suppressed the activation. The tBHQ-induced ARE binding activity was completely abrogated by an oxidizing agent, diamide, whereas 2-mercaptoethanol (2-ME) reversibly recovered the inhibitory effects of diamide, suggesting that ARE binding activity is redox-dependent. Moreover, overexpression of thioredoxin enhanced the ARE-mediated thioredoxin gene activation by tBHQ. Therefore, ARE-mediated induction of thioredoxin expression is a mechanism of enhancing signal transduction through the ARE in electrophile-induced stress responses.


Assuntos
Antioxidantes/metabolismo , Tiorredoxinas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Humanos , Hidroquinonas/farmacologia , Células K562 , Fator de Transcrição MafK , Fator 2 Relacionado a NF-E2 , Proteínas Nucleares/metabolismo , Oxirredução , Regiões Promotoras Genéticas , Tiorredoxinas/genética , Transativadores/metabolismo
12.
J Neurosci ; 23(2): 503-9, 2003 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-12533610

RESUMO

Thioredoxin (TRX) has a role in a variety of biological processes, including cytoprotection and the activation of transcription factors. Nerve growth factor (NGF) is a major survival factor of sympathetic neurons and promotes neurite outgrowth in rat pheochromocytoma PC12 cells. In this study, we showed that NGF induces TRX expression at protein and mRNA levels. NGF activated the TRX gene through a regulatory region positioned from -263 to -217 bp, containing the cAMP-responsive element (CRE). Insertion of a mutation in the CRE in this region abolished the response to NGF. NGF induced binding of CRE-binding protein to the CRE of the TRX promoter in an electrophoretic mobility shift assay. NGF also induced nuclear translocation of TRX. 2'-Amino-3'-methoxyflavone, an inhibitor of mitogen-activated protein kinase kinase, which is a known inhibitor of NGF-dependent differentiation in PC12 cells, suppressed the NGF-dependent expression and nuclear translocation of TRX. Overexpression of mutant TRX (32S/35S) or TRX antisense vector blocked the neurite outgrowth of PC12 cells by NGF. Overexpression of mutant TRX (C32S/C35S) suppressed the NGF-dependent activation of the CRE-mediated c-fos reporter gene. These results suggest that TRX plays a critical regulatory role in NGF-mediated signal transduction and outgrowth in PC12 cells.


Assuntos
Fator de Crescimento Neural/farmacologia , Neuritos/efeitos dos fármacos , Transdução de Sinais/fisiologia , Tiorredoxinas/metabolismo , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Substituição de Aminoácidos , Animais , Inibidores Enzimáticos/farmacologia , Expressão Gênica/efeitos dos fármacos , Genes Reporter , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Neuritos/fisiologia , Oligonucleotídeos Antissenso/biossíntese , Oligonucleotídeos Antissenso/farmacologia , Oxirredução , Células PC12 , Regiões Promotoras Genéticas/efeitos dos fármacos , Regiões Promotoras Genéticas/fisiologia , Proteínas Proto-Oncogênicas c-fos/biossíntese , RNA Mensageiro/metabolismo , Ratos , Elementos de Resposta/efeitos dos fármacos , Elementos de Resposta/fisiologia , Transdução de Sinais/efeitos dos fármacos , Tiorredoxinas/antagonistas & inibidores , Tiorredoxinas/genética
13.
Antioxid Redox Signal ; 6(1): 81-7, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-14713338

RESUMO

Thioredoxin (TRX) is induced by a variety of oxidative stimuli and shows cytoprotective roles against oxidative stress. To clarify the possibility of clinical application, we examined the effects of intravenously administered TRX in a model of transient focal cerebral ischemia in this study. Mature male C57BL/6j mice received either continuous intravenous infusion of recombinant human TRX (rhTRX) over a range of 1-10 mg/kg, bovine serum albumin, or vehicle alone for 2 h after 90-min transient middle cerebral artery occlusion (MCAO). Twenty-four hours after the transient MCAO, the animals were evaluated neurologically and the infarct volumes were assessed. Infarct volume, neurological deficit, and protein carbonyl contents, a marker of protein oxidation, in the brain were significantly ameliorated in rhTRX-treated mice at the dose of 3 and 10 mg/kg versus these parameters in control animals. Moreover, activation of p38 mitogen-activated protein kinase, whose pathway is involved in ischemic neuronal death, was suppressed in the rhTRX-treated mice. Further, rhTRX was detected in the ischemic hemisphere by western blot analysis, suggesting that rhTRX was able to permeate the blood-brain barrier in the ischemic hemisphere. These data indicate that exogenous TRX exerts distinct cytoprotective effects on cerebral ischemia/reperfusion injury in mice by means of its redox-regulating activity.


Assuntos
Infarto Encefálico/prevenção & controle , Encéfalo/efeitos dos fármacos , Ataque Isquêmico Transitório/tratamento farmacológico , Tiorredoxinas/uso terapêutico , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Infarto Encefálico/patologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Humanos , Injeções Intravenosas , Ataque Isquêmico Transitório/metabolismo , Ataque Isquêmico Transitório/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oxirredução/efeitos dos fármacos , Proteínas/metabolismo , Proteínas Recombinantes/uso terapêutico , Proteínas Quinases p38 Ativadas por Mitógeno
14.
Antioxid Redox Signal ; 4(4): 693-6, 2002 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12230882

RESUMO

Transgenic (Tg) mice overexpressing human thioredoxin (TRX), a small redox-active protein, were produced to investigate the role of the protein in a variety of stresses. Bone marrow cells from TRX-Tg mice were more resistant to ultraviolet C-induced cytocide compared with those from wild type (WT) C57BL/6 mice. TRX-Tg mice exhibited extended median and maximum life spans compared with WT mice. Telomerase activity in spleen tissues in TRX-Tg mice was higher than that in WT mice. These results suggest that overexpression of TRX results in resistance against oxidative stress and a possible extension of life span without apparent abnormality in mammals.


Assuntos
Longevidade , Estresse Oxidativo , Tiorredoxinas/genética , Tiorredoxinas/metabolismo , Animais , Células da Medula Óssea/metabolismo , Células da Medula Óssea/efeitos da radiação , Encéfalo/metabolismo , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miocárdio/metabolismo , Baço/enzimologia , Taxa de Sobrevida , Telomerase/metabolismo , Raios Ultravioleta
15.
Immunol Lett ; 92(1-2): 143-7, 2004 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-15081538

RESUMO

Thioredoxin (TRX) superfamily proteins that contain a conserved redox-active site -Cys-Xa.a.-Xa.a.-Cys- includes proinflammatory cytokine, macrophage migration inhibiting factor (MIF) and the immune regulatory cytokine, glycosylation inhibiting factor (GIF) in which Cys-60 is cysteinylated. In this report, we have analyzed the functional interaction between TRX and MIF/GIF. The stable Jurkat T cell line transfected with human TRX gene (TRX-transfectant) was highly resistant to hydrogen peroxide-induced apoptosis, but not the cell line transfected with vector (mock-transfectant). The expression level of MIF/GIF protein of TRX-transfectant was lower than that of mock-transfectant. Conversely, the expression level of intracellular TRX protein in CD4(+)-T cells derived from MIF -/- mice were significantly higher than that from background BALB/c mice. These findings collectively suggest that oxidative stress-induced apoptosis on T lymphocytes might be protected by the reciprocal regulation of TRX and MIF/GIF expression.


Assuntos
Cisteína/metabolismo , Linfocinas/imunologia , Fatores Inibidores da Migração de Macrófagos/imunologia , Tiorredoxinas/imunologia , Animais , Apoptose/fisiologia , Humanos , Peróxido de Hidrogênio/metabolismo , Oxirredutases Intramoleculares , Células Jurkat , Linfocinas/genética , Linfocinas/metabolismo , Fatores Inibidores da Migração de Macrófagos/genética , Fatores Inibidores da Migração de Macrófagos/metabolismo , Camundongos , Família Multigênica/imunologia , Família Multigênica/fisiologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Tiorredoxinas/metabolismo
16.
Redox Rep ; 7(5): 256-9, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12688505

RESUMO

Thioredoxin (TRX) is a 13 kDa protein with antioxidant effect and redox regulating functions. Peroxynitrite is a strong oxidizing and nitrating agent which can react with all classes of biomolecules. In the present study, we focused on the association between TRX and nitrotyrosine, which served as a marker of peroxynitrite formation, in the neonatal hypoxia-ischemia (HI) rat brain. At 4-16 h after HI, the immunoreactivity for TRX was diminished in the injured region in the cortex and striatum, whereas nitrotyrosine immunoreactivity was enhanced. In contrast, around the injured region, TRX immunoreactivity was enhanced in survival neurons at 4-24 h after HI, while the immunoreactivity for nitrotyrosine was mostly not detected. Northern blot analysis showed increased TRX mRNA induction in the cerebral hemisphere ipsilateral to the carotid ligation from 4-24 h after HI but not in the contralateral hypoxic hemisphere. These findings suggest that production of peroxynitrite is involved in HI brain injury, and that induced TRX plays a neuroprotective role against oxidative stress resulting from HI.


Assuntos
Encéfalo/patologia , Hipóxia , Traumatismo por Reperfusão , Tiorredoxinas/biossíntese , Tirosina/análogos & derivados , Tirosina/química , Animais , Animais Recém-Nascidos , Northern Blotting , Encéfalo/metabolismo , Modelos Anatômicos , Oxidantes/farmacologia , Ácido Peroxinitroso/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Tirosina/metabolismo
17.
PLoS One ; 9(4): e95080, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24762746

RESUMO

BACKGROUND AND AIMS: Azathioprine (AZA) is widely used for the treatment of inflammatory bowel disease (IBD) patients. AZA is catabolized by thiopurine S-methyltransferase (TPMT), which exhibits genetic polymorphisms. It has also been reported that 5-aminosalicylic acid (5-ASA) inhibits TPMT activity, and that increased 6-thioguanine nucleotide (6-TGN, a metabolite of AZA) blood concentrations result in an increased number of ADRs. In this study, single nucleotide polymorphisms (SNPs) related to differential gene expression affecting AZA drug metabolism in combination therapy with 5-ASA were examined. METHODS: To identify genetic biomarkers for the prediction of 6-TGN blood concentration, ExpressGenotyping analysis was used. ExpressGenotyping analysis is able to detect critical pharmacogenetic SNPs by analyzing drug-induced expression allelic imbalance (EAI) of premature RNA in HapMap lymphocytes. We collected blood samples on 38 patients with inflammatory bowel disease treated with AZA and corroboration of the obtained SNPs was attempted in clinical samples. RESULTS: A large number of SNPs with AZA/5-ASA-induced EAI within the investigated HapMap lymphocytes was identified by ExpressGenotyping analysis. The respective SNPs were analyzed in IBD patients' blood samples. Among these SNPs, several that have not yet been described to be induced by AZA/5-ASA were found. SNPs within SLC38A9 showed a particular correlation with patients' 6-TGN blood concentrations. CONCLUSIONS: Based on these results, ExpressGenotyping analysis and genotyping of patients appears to be a useful way to identify inter-individual differences in drug responses and ADRs to AZA/5-ASA. This study provides helpful information on genetic biomarkers for optimized AZA/5-ASA treatment of IBD patients.


Assuntos
Sistemas de Transporte de Aminoácidos/genética , Azatioprina/farmacocinética , Imunossupressores/farmacocinética , Doenças Inflamatórias Intestinais/sangue , Mesalamina/uso terapêutico , Adulto , Anti-Inflamatórios não Esteroides/uso terapêutico , Azatioprina/uso terapêutico , Células Cultivadas , Quimioterapia Combinada , Feminino , Estudos de Associação Genética , Marcadores Genéticos , Humanos , Imunossupressores/uso terapêutico , Doenças Inflamatórias Intestinais/tratamento farmacológico , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único
18.
Invest Ophthalmol Vis Sci ; 49(11): 5118-25, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18515569

RESUMO

PURPOSE: To examine the role of thioredoxin-1 (TRX-1), an endogenous protein with a variety of redox-related roles, in the formation of choroidal neovascularization (CNV). METHODS: CNV was induced by laser photocoagulation of the ocular fundus in wild-type and transgenic mice overexpressing human TRX-1 (TRX-1 Tg). Mice were injected intraperitoneally with TRX-1, mutant TRX, or vehicle. The incidence of CNV was evaluated by lectin staining. Leukocyte recruitment and C3b deposition after laser injury were determined by immunohistochemistry and Western blotting. Moreover, TRX-1-associated proteins from human plasma were isolated by two-dimensional gel electrophoresis with the use of a column coupled with a mutant TRX-1 and were identified by mass spectrometry and proteomics analysis. Complement activation was determined by a fluid-phase RESULTS: The incidence of laser-induced CNV was reduced in TRX-1 Tg mice (56.1%) and in C57B/6 mice treated with TRX-1 (46.7%) but not in mutant TRX-1 (79.2%) compared with wild-type mice (85.7%). Furthermore, leukocyte recruitment was prevented in TRX-1-treated mice; C3b deposition was decreased in these and TRX-1 Tg mice. In human plasma, five proteins associated with TRX-1 were identified as apolipoprotein A-I, the CD5 antigen-like member of the scavenger receptor, cysteine-rich superfamily fibrinogen, albumin, and complement factor H (CFH). TRX-1 inhibited the alternative pathway C3 convertase, and its effect was additive with CFH. CONCLUSIONS: These findings show that TRX-1 interacts with CFH, regulates complement activity, and inhibits CNV, suggesting novel preventive and interventional therapeutic strategies for AMD.


Assuntos
Neovascularização de Coroide/prevenção & controle , Fator H do Complemento/metabolismo , Tiorredoxinas/uso terapêutico , Animais , Western Blotting , Neovascularização de Coroide/metabolismo , Neovascularização de Coroide/patologia , Fator H do Complemento/efeitos dos fármacos , Modelos Animais de Doenças , Eletroforese em Gel Bidimensional , Humanos , Imuno-Histoquímica , Injeções Intraperitoneais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Tiorredoxinas/administração & dosagem
19.
Eur J Immunol ; 38(5): 1358-67, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18425727

RESUMO

Thioredoxin-binding protein-2 (TBP-2), also known as vitamin D3-up-regulated protein 1 (VDUP1), was identified as an endogenous molecule interacting with thioredoxin (TRX). Here, we show that dendritic cells (DC) derived from TBP-2-deficient mice are defective in the function of T cell activation. To compare TBP-2(-/-) DC function with wild-type (WT) DC, we stimulated DC with lipopolysaccharide (LPS). Although TBP-2(-/-) DC and WT DC expressed comparable levels of MHC class II and costimulatory molecules such as CD40, CD80 and CD86, the IL-12p40, IL-12p70 and IL-6 productions of TBP-2(-/-) DC were attenuated. In a mixed leukocyte reaction (MLR), the concentrations of IL-2, IFN-gamma, IL-4 and IL-10 in the culture supernatant of MLR with TBP-2(-/-) DC were significantly lower than those in the cultures with WT DC. In MLR also, as with LPS stimulation, IL-12p40 and IL-12p70 production from TBP-2(-/-) DC was less than that from WT DC. Proliferation of T cells cultured with TBP-2(-/-) DC was poorer than that with WT DC. In vivo delayed-type hypersensitivity responses in TBP-2(-/-) mice immunized with ovalbumin were significantly reduced compared to WT mice. These results indicate that TBP-2 plays a crucial role in DC to induce T cell responses.


Assuntos
Proteínas de Transporte/fisiologia , Células Dendríticas/fisiologia , Linfócitos T/imunologia , Tiorredoxinas/fisiologia , Animais , Formação de Anticorpos/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Proliferação de Células , Citocinas/genética , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Feminino , Expressão Gênica , Hipersensibilidade Tardia/induzido quimicamente , Hipersensibilidade Tardia/genética , Hipersensibilidade Tardia/imunologia , Interleucina-12/genética , Interleucina-12/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Ativação Linfocitária , Teste de Cultura Mista de Linfócitos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Ovalbumina/imunologia , Linfócitos T/metabolismo
20.
J Neurochem ; 98(2): 372-85, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16805832

RESUMO

Thioredoxin (TRX) plays a variety of redox-related roles in organisms. To investigate its function as an endogenous redox regulator in NMDA-induced retinal neurotoxicity, we injected NMDA with TRX, mutant TRX or saline into the vitreous cavity of rat eyes. Retinal ganglion cells were rescued by TRX, compared with saline, when evaluated by retrograde labeling analysis at 7 days after NMDA injection. TRX, but not its mutant form, prevented NMDA-induced apoptosis in the retina, as measured by terminal deoxynucleotidyl transferase-mediated UTP nick-end labeling. The induction of caspase 3 and 9, but not caspase 8, by NMDA was significantly lower in TRX-treated eyes than in saline-treated eyes. NMDA-induced activation of the MAPKs, p38 kinase and c-Jun N-terminal kinase after 6 h and of the MAPK kinases (MKKs) MKK3/6 and MKK4 after 3 h was markedly suppressed in retinal ganglion cells by TRX but not by the mutant form. NMDA-induced increases in protein carbonylation, nitrosylation and lipid peroxidation were also suppressed in TRX-treated eyes. We concluded that the intravitreous injection of TRX effectively attenuated NMDA-induced retinal cell damage and that suppression of oxidative stress and inhibition of apoptotic signaling pathways were involved in this neuroprotection.


Assuntos
Agonistas de Aminoácidos Excitatórios/farmacologia , N-Metilaspartato/antagonistas & inibidores , Fármacos Neuroprotetores , Doenças Retinianas/induzido quimicamente , Doenças Retinianas/prevenção & controle , Tiorredoxinas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Caspase 3 , Caspase 8 , Caspase 9 , Caspases/metabolismo , Agonistas de Aminoácidos Excitatórios/toxicidade , Técnicas Imunoenzimáticas , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Proteínas Quinases Ativadas por Mitógeno/metabolismo , N-Metilaspartato/toxicidade , Fosforilação , Proteínas Proto-Oncogênicas c-jun/metabolismo , Ratos , Ratos Wistar , Retina/patologia , Doenças Retinianas/patologia , Células Ganglionares da Retina/efeitos dos fármacos , Tirosina/análogos & derivados , Tirosina/metabolismo
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