RESUMO
cis-Aconitic acid is a constituent from the leaves of Echinodorus grandiflorus, a medicinal plant traditionally used in Brazil to treat inflammatory conditions, including arthritic diseases. The present study aimed to investigate the anti-arthritic effect of cis-aconitic acid in murine models of antigen-induced arthritis and monosodium urate-induced gout. The possible underlying mechanisms of action was evaluated in THP-1 macrophages. Oral treatment with cis-aconitic acid (10, 30, and 90 mg/kg) reduced leukocyte accumulation in the joint cavity and C-X-C motif chemokine ligand 1 and IL-1ß levels in periarticular tissue. cis-Aconitic acid treatment reduced joint inflammation in tissue sections of antigen-induced arthritis mice and these effects were associated with decreased mechanical hypernociception. Administration of cis-aconitic acid (30 mg/kg p.âo.) also reduced leukocyte accumulation in the joint cavity after the injection of monosodium urate crystals. cis-Aconitic acid reduced in vitro the release of TNF-α and phosphorylation of IκBα in lipopolysaccharide-stimulated THP-1 macrophages, suggesting that inhibition of nuclear factor kappa B activation was an underlying mechanism of cis-aconitic acid-induced anti-inflammatory effects. In conclusion, cis-aconitic acid has significant anti-inflammatory effects in antigen-induced arthritis and monosodium urate-induced arthritis in mice, suggesting its potential for the treatment of inflammatory diseases of the joint in humans. Additionally, our findings suggest that this compound may contribute to the anti-inflammatory effect previously reported for E. grandiflorus extracts.
Assuntos
Alismataceae , Gota , Humanos , Camundongos , Animais , Ácido Aconítico/farmacologia , Inibidor de NF-kappaB alfa , Ácido Úrico , Lipopolissacarídeos , NF-kappa B , Fator de Necrose Tumoral alfa , Ligantes , Alismataceae/química , Gota/induzido quimicamente , Gota/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Quimiocinas , InflamaçãoRESUMO
This study investigates the participation of PI3Kγ in the development of joint inflammation and dysfunction in an experimental model of acute gout in mice. Acute gout was induced by injection of monosodium urate (MSU) crystals into the tibiofemoral joint of mice. The involvement of PI3Kγ was evaluated using a selective inhibitor and mice deficient for PI3Kγ (PI3Kγ-/- ) or with loss of kinase activity. Neutrophils recovered from the inflamed joint were quantified and stained for phosphorylated Akt (pAkt) and production of reactive oxygen species (ROS). The adherence of leukocytes to the joint microvasculature was assessed by intravital microscopy and cleaved caspase-1 by Western blot. Injection of MSU crystals induced massive accumulation of neutrophils expressing phosphorylated Akt. In the absence of PI3Kγ, there was reduction of pAkt expression, chemokine production, and neutrophil recruitment. Genetic or pharmacological inhibition of PI3Kγ reduced the adherence of leukocytes to the joint microvasculature, even in joints with established inflammation. Neutrophils from PI3Kγ-/- mice produced less ROS than wild-type neutrophils. There was decreased joint damage and dysfunction in the absence of PI3Kγ. In addition, in the absence of PI3Kγ activity, there was reduction of cleaved caspase-1 and IL-1ß production in synovial tissue after injection of MSU crystals and leukotriene B4 . Our studies suggest that PI3Kγ is crucial for MSU crystal-induced acute joint inflammation. It is necessary for regulating caspase-1 activation and for mediating neutrophil migration and activation. Drugs that impair PI3Kγ function may be useful to control acute gout inflammation.
Assuntos
Artrite Gotosa/enzimologia , Artrite Gotosa/imunologia , Caspase 1/metabolismo , Classe Ib de Fosfatidilinositol 3-Quinase/metabolismo , Infiltração de Neutrófilos , Doença Aguda , Animais , Adesão Celular , Movimento Celular , Classe Ib de Fosfatidilinositol 3-Quinase/deficiência , Citoplasma/metabolismo , Ativação Enzimática , Inflamassomos/metabolismo , Inflamação/patologia , Interleucina-1beta/metabolismo , Articulações/patologia , Leucotrieno B4/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Microvasos/patologia , Neutrófilos/metabolismo , Nociceptividade , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Membrana Sinovial/irrigação sanguínea , Ácido ÚricoRESUMO
The leaves of Echinodorus grandiflorus are traditionally used in Brazil to treat several inflammatory conditions, including arthritis. This study aimed to investigate the antiarthritis activity of the 70% ethanol extract of E. grandiflorus leaves and a standardized flavonoid-rich fraction in an antigen-induced arthritis model in mice. Previously immunized mice were treated per os with saline (control group), 70% ethanol extract (100-1000 mg/kg), or a flavonoid-rich fraction (0.7-7.2 mg/kg) 40 minutes before and 3 and 6 hours after the challenge with antigen into the knee joint. The administration of the 70% ethanol extract and flavonoid-rich fraction to mice significantly reduced neutrophil recruitment to the joint cavity and in periarticular tissue. The levels of chemokine (C-X-C motif) ligand 1, tumor necrosis factor-α, and interleukin-1ß quantified by the enzyme-linked immunosorbent assay (ELISA) in the periarticular tissue were also diminished in mice treated with the 70% ethanol extract and flavonoid-rich fraction, as well as mechanical hypernociception. Histological analysis confirmed that both the 70% ethanol extract and flavonoid-rich fraction suppressed joint inflammation and inhibited cartilage and bone destruction when compared to the control group. Our results demonstrate, for the first time, that E. grandiflorus has anti-inflammatory activity in an experimental arthritis model and highlights the role of flavonoids in the observed response.
Assuntos
Alismataceae/química , Anti-Inflamatórios não Esteroides/uso terapêutico , Artrite Experimental/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Animais , Brasil , Modelos Animais de Doenças , Flavonoides/uso terapêutico , Glicosídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Monossacarídeos/uso terapêutico , Folhas de Planta/químicaRESUMO
Angiotensin (Ang) II and its AT1 receptors have been implicated in the pathogenesis of rheumatoid arthritis. Activation of the counter-regulatory Ang-(1-7)-Mas receptor axis may contribute to some of the effects of AT1 receptor blockers (ARBs). In this study, we have used losartan, an ARB, to investigate the role of and the mechanisms by which AT1 receptors participated in two experimental models of arthritis: antigen-induced arthritis (AIA) in mice and adjuvant-induced arthritis (AdIA) in rats. Treatment with losartan decreased neutrophil recruitment, hypernociception and the production of TNF-α, IL-1ß and chemokine (C-X-C motif) ligand 1 in mice subjected to AIA. Histopathological analysis showed significant reduction of tissue injury and inflammation and decreased proteoglycan loss. In addition to decreasing cytokine production, losartan directly reduced leukocyte rolling and adhesion. Anti-inflammatory effects of losartan were not associated to Mas receptor activation and/or Ang-(1-7) production. Anti-inflammatory effects were reproduced in rats subjected to AdIA. This study shows that ARBs have potent anti-inflammatory effects in animal models of arthritis. Mechanistically, reduction of leukocyte accumulation and of joint damage was associated with local inhibition of cytokine production and direct inhibition of leukocyte-endothelium interactions. The anti-inflammatory actions of losartan were accompanied by functional improvement of the joint, as seen by reduced joint hypernociception. These findings support the use of ARBs for the treatment of human arthritis and provide potential mechanisms for the anti-inflammatory actions of these compounds.
Assuntos
Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Artrite Experimental/tratamento farmacológico , Losartan/farmacologia , Receptor Tipo 1 de Angiotensina/efeitos dos fármacos , Angiotensina I/biossíntese , Animais , Artrite Reumatoide/tratamento farmacológico , Adesão Celular/efeitos dos fármacos , Quimiocina CXCL1/biossíntese , Modelos Animais de Doenças , Feminino , Hiperalgesia/tratamento farmacológico , Inflamação/tratamento farmacológico , Interleucina-1beta/biossíntese , Migração e Rolagem de Leucócitos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Infiltração de Neutrófilos/efeitos dos fármacos , Fragmentos de Peptídeos/biossíntese , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
OBJECTIVES: To investigate the effect of 2 standardized exercise programs, muscle strength exercises (SE) and aerobic exercises (AE), on the plasma levels of brain-derived neurotrophic factor (BDNF) and depressive symptoms in 451 elderly women. DESIGN: A randomized controlled trial. SETTING: Belo Horizonte/MG-Brazil. PARTICIPANTS: Community-dwelling older women (N=451; age, 65-89y). INTERVENTION: The participants were divided into 2 groups: SE and AE. Both protocols lasted 10 weeks, and 30 sessions (1-h sessions) in total were performed 3 times a week under the direct supervision of physical therapists. MAIN OUTCOME MEASURES: Plasma levels of BDNF (enzyme-linked immunosorbent assay) and depressive symptoms (Geriatric Depression Scale). RESULTS: There was a significant difference for BDNF plasma levels between the SE and AE groups (P=.009). Post hoc analysis revealed a pre-post intervention difference in BDNF levels only for the SE group (P=.008). A statistically significant difference was found for the pre- and postintervention Geriatric Depression Scale scores in both groups (P=.001), showing that the effects of both exercise protocols were comparable regarding depressive symptoms (P=.185). CONCLUSIONS: The present findings have demonstrated the positive effect of muscle strengthening and aerobic intervention on depressive symptoms in community-dwelling elderly women. Interestingly, only SE significantly increased the plasma levels of BDNF in our sample. The positive effects of physical exercise on depressive symptoms in the elderly were not mediated by BDNF.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/sangue , Transtorno Depressivo/sangue , Transtorno Depressivo/reabilitação , Exercício Físico , Fatores Etários , Idoso , Brasil , Estudos de Coortes , Feminino , Humanos , Fatores SexuaisRESUMO
OBJECTIVE: Deposition of monosodium urate monohydrate (MSU) crystals in the joints promotes an intense inflammatory response and joint dysfunction. This study evaluated the role of the NLRP3 inflammasome and 5-lipoxygenase (5-LOX)-derived leukotriene B(4) (LTB(4) ) in driving tissue inflammation and hypernociception in a murine model of gout. METHODS: Gout was induced by injecting MSU crystals into the joints of mice. Wild-type mice and mice deficient in NLRP3, ASC, caspase 1, interleukin-1ß (IL-1ß), IL-1 receptor type I (IL-1RI), IL-18R, myeloid differentiation factor 88 (MyD88), or 5-LOX were used. Evaluations were performed to assess neutrophil influx, LTB(4) activity, cytokine (IL-1ß, CXCL1) production (by enzyme-linked immunosorbent assay), synovial microvasculature cell adhesion (by intravital microscopy), and hypernociception. Cleaved caspase 1 and production of reactive oxygen species (ROS) were analyzed in macrophages by Western blotting and fluorometric assay, respectively. RESULTS: Injection of MSU crystals into the knee joints of mice induced neutrophil influx and neutrophil-dependent hypernociception. MSU crystal-induced neutrophil influx was CXCR2-dependent and relied on the induction of CXCL1 in an NLRP3/ASC/caspase 1/IL-1ß/MyD88-dependent manner. LTB(4) was produced rapidly after injection of MSU crystals, and this was necessary for caspase 1-dependent IL-1ß production and consequent release of CXCR2-acting chemokines in vivo. In vitro, macrophages produced LTB(4) after MSU crystal injection, and LTB(4) was relevant in the MSU crystal-induced maturation of IL-1ß. Mechanistically, LTB(4) drove MSU crystal-induced production of ROS and ROS-dependent activation of the NLRP3 inflammasome. CONCLUSION: These results reveal the role of the NLRP3 inflammasome in mediating MSU crystal-induced inflammation and dysfunction of the joints, and highlight a previously unrecognized role of LTB(4) in driving NLRP3 inflammasome activation in response to MSU crystals, both in vitro and in vivo.
Assuntos
Proteínas de Transporte/metabolismo , Gota/metabolismo , Hiperalgesia/metabolismo , Inflamassomos/metabolismo , Leucotrieno B4/metabolismo , Infiltração de Neutrófilos/fisiologia , Neutrófilos/metabolismo , Animais , Caspase 1/metabolismo , Citocinas/metabolismo , Gota/induzido quimicamente , Gota/imunologia , Hiperalgesia/imunologia , Inflamassomos/imunologia , Inflamação/imunologia , Inflamação/metabolismo , Interleucina-1beta/metabolismo , Leucotrieno B4/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Knockout , Proteína 3 que Contém Domínio de Pirina da Família NLR , Neutrófilos/imunologia , Espécies Reativas de Oxigênio/metabolismo , Membrana Sinovial/imunologia , Membrana Sinovial/metabolismo , Ácido Úrico/farmacologiaRESUMO
BACKGROUND: Frailty syndrome in elderly people is characterized by a reduction of energy reserves and also by a decreased of resistance to stressors, resulting in an increase of vulnerability. OBJECTIVE: The aim of this study was to verify the effect of a muscle-strengthening program with load in pre-frail elder women with regards to the functional capacity, knee extensor muscle strength and their correlation. METHODS: Thrity-two pre-frail community-dwelling women participated in this study. Potential participants with cognitive impairment (MEEM), lower extremities orthopedic surgery, fractures, inability to walk unaided, neurological diseases, acute inflammatory disease, tumor growth, regular physical activity and current use of immunomodulators were excluded. All partcipants were evaluated by a blinded assessor using: Timed up and go (TUG), 10-Meter Walk Test (10MWT) and knee extensor muscle strength (Byodex System 3 Pro® isokinetic dynamometer at angular speeds of 60 and 180(0)/s). The intervention consisted of strengthening exercises of the lower extremities at 70% of 1RM, three times/ week for ten weeks. The statistical analysis was performed using the ANOVA and Spearman tests RESULTS: After the intervention, it was observed statistical significance on the work at 180(0)/s (F=12.71, p=0.02), on the power at 180(0)/s (F=15.40, p=0.02) and on the functional capacity (TUG, F=9.54, p=0.01; TC10, F=3.80, p=0.01). There was a good negative and statistically significant correlation between the TUG and work at 60(0)/s, such as the TUG and work at 180(0)/s (r=-0.65, p=0.01; r=-0.72, p=0.01). CONCLUSION: The intervention improved the muscular power and the functional capacity. The increase of the power correlated with function, which is an important variable of the quality of life in the pre-frail elders. Article registered in the ISRCT register under number ISRCTN62824599.
Assuntos
Terapia por Exercício , Força Muscular , Idoso , Estudos Cross-Over , Feminino , Idoso Fragilizado , Humanos , Joelho/fisiologia , Características de Residência , Método Simples-CegoRESUMO
CONTEXTUALIZAÇÃO: Na síndrome de fragilidade em idosos, há diminuição das reservas de energia e resistência aos estressores, com aumento da vulnerabilidade. OBJETIVO: Verificar o efeito do treinamento de força muscular com carga na capacidade funcional e força muscular dos extensores do joelho e sua associação, após treinamento, em idosas pré-frágeis da comunidade. MÉTODOS:Participaram 32 idosas, pré-frágeis, da comunidade. Excluíram-se aquelas com Miniexame do Estado Mental (MEEM) incompatível; cirurgias ortopédicas dos membros inferiores; fraturas; doenças neurológicas; doenças inflamatórias agudas; neoplasias; atividade física regular; uso de medicamento com ação no sistema imunológico e sem marcha independente. Avaliou-se a capacidade funcional (Timed Up and Go - TUG e velocidade de marcha - TC10) e a força muscular dos extensores do joelho (Byodex System 3 Pro®) nas velocidades angulares de 60 e 180(0)/s. Para o fortalecimento muscular, utilizou-se carga de 75 por cento de resistência máxima (1RM), durante dez semanas, três vezes/semana. A análise estatística foi feita pela ANOVA e Spearman (α=5 por cento). RESULTADOS: Após o treinamento, houve melhora estatística do trabalho normalizado em 180(0)/s (F=12,71, p=0,02), na potência, em 180(0)/s (F=15,40, p=0,02) e na capacidade funcional (TUG, F=9,54, p=0,01; TC10, F=3,80, p=0,01). Houve boa correlação negativa significativa do TUG com as medidas de trabalho normalizado em 60 e 180(0)/s (r=-0,65, p=0,01; r=-0,72, p=0,01). CONCLUSÃO: O treinamento produziu melhora da potência muscular e capacidade funcional. A melhora da potência associou-se à melhora funcional, importante variável para a qualidade de vida de idosas pré-frágeis. Artigo registrado no ISRCT register sob o número ISRCTN62824599.
BACKGROUND: Frailty syndrome in elderly people is characterized by a reduction of energy reserves and also by a decreased of resistance to stressors, resulting in an increase of vulnerability. OBJECTIVE: The aim of this study was to verify the effect of a muscle-strengthening program with load in pre-frail elder women with regards to the functional capacity, knee extensor muscle strength and their correlation. METHODS: Thrity-two pre-frail community-dwelling women participated in this study. Potential participants with cognitive impairment (MEEM), lower extremities orthopedic surgery, fractures, inability to walk unaided, neurological diseases, acute inflammatory disease, tumor growth, regular physical activity and current use of immunomodulators were excluded. All partcipants were evaluated by a blinded assessor using: Timed up and go (TUG), 10-Meter Walk Test (10MWT) and knee extensor muscle strength (Byodex System 3 Pro® isokinetic dynamometer at angular speeds of 60 and 180(0)/s). The intervention consisted of strengthening exercises of the lower extremities at 70 percent of 1RM, three times/ week for ten weeks. The statistical analysis was performed using the ANOVA and Spearman tests RESULTS: After the intervention, it was observed statistical significance on the work at 180(0)/s (F=12.71, p=0.02), on the power at 180(0)/s (F=15.40, p=0.02) and on the functional capacity (TUG, F=9.54, p=0.01; TC10, F=3.80, p=0.01). There was a good negative and statistically significant correlation between the TUG and work at 60(0)/s, such as the TUG and work at 180(0)/s (r=-0.65, p=0.01; r=-0.72, p=0.01). CONCLUSION: The intervention improved the muscular power and the functional capacity. The increase of the power correlated with function, which is an important variable of the quality of life in the pre-frail elders. Article registered in the ISRCT register under number ISRCTN62824599.
Assuntos
Idoso , Feminino , Humanos , Terapia por Exercício , Força Muscular , Estudos Cross-Over , Idoso Fragilizado , Joelho/fisiologia , Características de Residência , Método Simples-CegoRESUMO
OBJECTIVE: Neutrophil accumulation contributes to the pathogenesis of rheumatoid arthritis. This study was undertaken to examine the ability of H2O2 to influence neutrophilic inflammation in a model of antigen-induced arthritis (AIA) in mice. METHODS: AIA was induced by administration of antigen into the knee joints of previously immunized mice. Neutrophil accumulation was measured by counting neutrophils in the synovial cavity and assaying myeloperoxidase activity in the tissue surrounding the mouse knee joint. Apoptosis was determined by morphologic and molecular techniques. The role of H2O2 was studied using mice that do not produce reactive oxygen species (gp91phox-/- mice) and drugs that enhance the generation or enhance the degradation of H2O2. RESULTS: Antigen challenge of immunized mice induced neutrophil accumulation that peaked at 12-24 hours after challenge. H2O2 production peaked at 24 hours, after which time, the inflammation resolved. Neutrophil recruitment was similar in wild-type and gp91phox-/- mice, but there was delayed resolution in gp91phox-/- mice or after administration of catalase. In contrast, administration of H2O2 or superoxide dismutase (SOD) resolved neutrophilic inflammation. The resolution of inflammation induced by SOD or H2O2 was accompanied by an increase in the number of apoptotic neutrophils. Apoptosis was associated with an increase in Bax and caspase 3 cleavage and was secondary to phosphatidylinositol 3-kinase (PI3K)/Akt activation. CONCLUSION: Our findings indicate that levels of H2O2 increase during neutrophil influx and are necessary for the natural resolution of neutrophilic inflammation. Mechanistically, enhanced levels of H2O2 (endogenous or exogenous) inhibit p-Akt/NF-κB and induce apoptosis of migrated neutrophils. Modulation of H2O2 production may represent a novel strategy for controlling neutrophilic inflammation in the joints.
Assuntos
Artrite Experimental/imunologia , Peróxido de Hidrogênio/metabolismo , Neutrófilos/imunologia , Membrana Sinovial/metabolismo , Animais , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/imunologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Articulações/imunologia , Articulações/metabolismo , Articulações/patologia , Masculino , Camundongos , Neutrófilos/metabolismo , Neutrófilos/patologia , Membrana Sinovial/imunologia , Membrana Sinovial/patologiaRESUMO
UNLABELLED: BACKGROUND AND PURPOSE; Chronic joint inflammation and pain are the hallmarks of disease in patients with inflammatory arthritis, notably rheumatoid arthritis. The aim of the present study was to investigate the relative contribution of tumour necrosis factor (TNF)-α, interleukin (IL)-1ß and neutrophil influx for joint inflammation and nociception in a novel murine model of antigen-induced arthritis (AIA). EXPERIMENTAL APPROACH: AIA was induced by administration of antigen into knee joint of previously immunized mice. Neutrophil accumulation was determined by counting neutrophils in the joints and assessing myeloperoxidase activity in tissues surrounding the joints. TNF-α, IL-1ß and CXCL-1 were measured by elisa. Mechanical hypernociception was assessed in parallel, using an electronic pressure meter. KEY RESULTS: Hypernociception was dependent on antigen dose and the time after its administration; it was prevented by treatment with morphine and associated with neutrophil infiltration and local production of TNF-α, IL-1ß and CXCL-1. Administration of a chimeric monoclonal antibody to TNF-α (infliximab) or IL-1receptor antagonist prevented neutrophil influx and hypernociception, and this was comparable to the effects of dexamethasone. Treatment with fucoidin (a leucocyte adhesion inhibitor) greatly suppressed neutrophil influx and local production of TNF-α and IL-1ß, and hypernociception. CONCLUSIONS AND IMPLICATIONS: In conclusion, the present study describes a new model that allows for the concomitant evaluation of articular hypernociception and inflammation. Using this system, we demonstrated that a positive feedback loop involving neutrophil influx and the pro-inflammatory cytokines TNF-α and IL-1ß is necessary for articular hypernociception after antigen challenge of immunized mice.