RESUMO
Claudins are transmembrane proteins constituting one of three tight junction protein families. In patients with inflammatory bowel disease (IBD), disease activity-dependent changes in expression of certain claudins have been noted, thus making certain claudin family members potential therapy targets. A study was undertaken with the aim of exploring expression of claudins in human disease and two different animal models of IBD: dextrane sulfate sodium-induced colitis and adoptive transfer model of colitis. The expression of sealing claudin-1, claudin-3, claudin-4, and claudin-8, and pore-forming claudin-2 in humans and rodents has been evaluated by immunohistochemistry and quantitative polymerase chain reaction. Claudins were expressed by epithelial and cells of mesodermal origin and were found to be situated at the membrane, within the cytoplasm, or within the nuclei. Claudin expression by human mononuclear cells isolated from lamina propria has been confirmed by Western blot and flow cytometry. The claudin expression pattern in uninflamed and inflamed colon varied between species and murine strains. In IBD and both animal models, diverse alterations in claudin expression by epithelial and inflammatory cells were recorded. Tissue mRNA levels for each studied claudin reflected changes within cell lineage and, at the same time, mirrored the ratio between various cell types. Based on the results of the study, it can be concluded that 1) claudins are not expressed exclusively by epithelial cells, but by certain types of cells of mesodermal origin as well; 2) changes in the claudin mRNA level should be interpreted in the context of overall tissue alterations; and 3) both IBD animal models that were analyzed can be used for investigating claudins as a therapy target, respecting their similarities and differences highlighted in this study.
RESUMO
Several antibiotics demonstrate both antibacterial and anti-inflammatory/immunomodulatory activities and are used to treat inflammatory pulmonary disorders. Lefamulin is a pleuromutilin antibiotic approved to treat community-acquired bacterial pneumonia (CABP). This study evaluated lefamulin anti-inflammatory effects in vivo and in vitro in a lipopolysaccharide-induced lung neutrophilia model in which mouse airways were challenged with intranasal lipopolysaccharide. Lefamulin and comparators azithromycin and dexamethasone were administered 30min before lipopolysaccharide challenge; neutrophil infiltration into BALF and inflammatory mediator induction in lung homogenates were measured 4h postchallenge. Single subcutaneous lefamulin doses (10â140mg/kg) resulted in dose-dependent reductions of BALF neutrophil cell counts, comparable to or more potent than subcutaneous azithromycin (10â100mg/kg) and oral/intraperitoneal dexamethasone (0.5/1mg/kg). Lipopolysaccharide-induced pro-inflammatory cytokine (TNF-α, IL-6, IL-1ß, and GM-CSF), chemokine (CXCL-1, CXCL-2, and CCL-2), and MMP-9 levels were significantly and dose-dependently reduced in mouse lung tissue with lefamulin; effects were comparable to or more potent than with dexamethasone or azithromycin. Pharmacokinetic analyses confirmed exposure-equivalence of 30mg/kg subcutaneous lefamulin in mice to a single clinical lefamulin dose to treat CABP in humans (150mg intravenous/600mg oral). In vitro, neither lefamulin nor azithromycin had any relevant influence on lipopolysaccharide-induced cytokine/chemokine levels in J774.2 mouse macrophage or human peripheral blood mononuclear cell supernatants, nor were any effects observed on IL-8âinduced human neutrophil chemotaxis. These in vitro results suggest that impediment of neutrophil infiltration by lefamulin in vivo may not occur through direct interaction with macrophages or neutrophilic chemotaxis. This is the first study to demonstrate inhibition of neutrophilic lung infiltration and reduction of pro-inflammatory cytokine/chemokine concentrations by clinically relevant lefamulin doses. This anti-inflammatory activity may be beneficial in patients with acute respiratory distress syndrome, cystic fibrosis, or severe inflammation-mediated lung injury, similar to glucocorticoid (eg, dexamethasone) activity. Future lefamulin anti-inflammatory/immunomodulatory activity studies are warranted to further elucidate mechanism of action and evaluate clinical implications.
Assuntos
Azitromicina , Diterpenos , Compostos Policíclicos , Tioglicolatos , Animais , Infecções Comunitárias Adquiridas , Leucócitos Mononucleares , Lipopolissacarídeos , CamundongosRESUMO
Autotaxin is a circulating enzyme with a major role in the production of lysophosphatic acid (LPA) species in blood. A role for the autotaxin/LPA axis has been suggested in many disease areas including pulmonary fibrosis. Structural modifications of the known autotaxin inhibitor lead compound 1, to attenuate hERG inhibition, remove CYP3A4 time-dependent inhibition, and improve pharmacokinetic properties, led to the identification of clinical candidate GLPG1690 (11). Compound 11 was able to cause a sustained reduction of LPA levels in plasma in vivo and was shown to be efficacious in a bleomycin-induced pulmonary fibrosis model in mice and in reducing extracellular matrix deposition in the lung while also reducing LPA 18:2 content in bronchoalveolar lavage fluid. Compound 11 is currently being evaluated in an exploratory phase 2a study in idiopathic pulmonary fibrosis patients.
Assuntos
Fibrose Pulmonar Idiopática/tratamento farmacológico , Imidazóis/uso terapêutico , Diester Fosfórico Hidrolases/efeitos dos fármacos , Pirimidinas/uso terapêutico , Animais , Humanos , Imidazóis/farmacologia , Camundongos , Camundongos Knockout , Diester Fosfórico Hidrolases/genética , Pirimidinas/farmacologia , RatosRESUMO
Although inhaled glucocorticoids are known to have systemic effects on bone metabolism, there is little comparative information on their relative potencies. The effects of three standard glucocorticoids in causing changes in bone metabolism and growth, therefore, were investigated in relation to other systemic effects in the rat. Given to male Sprague-Dawley rats, 4.5-5.5 weeks old, subcutaneously (s.c.), at doses of 0.3-10 mg/kg daily for 7 days, beclomethasone dipropionate, prednisolone and ciclesonide all dose-dependently inhibited thymus body mass index (BMI) (by 57%, 44% and 76% at 3 mg/kg). Ciclesonide, potently and prednisolone, less effectively, also repressed femoral bone growth (by 41% and 18% at 10 mg/kg), significantly reducing body weight gain (both by 100% at 10 mg/kg), and serum concentrations of acid phosphatase (ACP) and tartarate resistant acid phosphatase (TRACP) (by >30% at 10 mg/kg); both increased serum glucose and triglycerides levels. Serum alkaline phosphatase (ALP) was not affected. Beclomethasone dipropionate had little or no effect on these additional variables. In conclusion, ciclesonide showed pronounced bone growth inhibiting activity after s.c. administration to the rat while other two glucocorticoids showed differences in activity on bone metabolism. However, this model is sufficiently sensitive and specific for testing the effect of glucocorticoids on bone metabolism.
Assuntos
Beclometasona/toxicidade , Desenvolvimento Ósseo/efeitos dos fármacos , Fêmur/efeitos dos fármacos , Glucocorticoides/toxicidade , Prednisolona/toxicidade , Pregnenodionas/toxicidade , Fosfatase Ácida/sangue , Animais , Biomarcadores/sangue , Relação Dose-Resposta a Droga , Fêmur/crescimento & desenvolvimento , Fêmur/metabolismo , Fêmur/patologia , Isoenzimas/sangue , Masculino , Tamanho do Órgão , Ratos Sprague-Dawley , Fosfatase Ácida Resistente a Tartarato , Timo/efeitos dos fármacos , Timo/patologia , Aumento de Peso/efeitos dos fármacosRESUMO
A new concept in design of safe glucocorticoid therapy was introduced by conjugating potent glucocorticoid steroids with macrolides (macrolactonolides). These compounds were synthesized from various steroid 17ß-carboxylic acids and 9a-N-(3-aminoalkyl) derivatives of 9-deokso-9a-aza-9a-homoeritromicin A and 3-descladinosyl-9-deokso-9a-aza-9a-homoeritromicin A using stable alkyl chain. Combining property of macrolides to preferentially accumulate in immune cells, especially in phagocyte cells, with anti-inflammatory activity of classic steroids, we designed molecules which showed good anti-inflammatory activity in ovalbumin (OVA) induced asthma in rats. The synthesis, in vitro and in vivo anti-inflammatory activity of this novel class of compounds are described.
Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/uso terapêutico , Asma/tratamento farmacológico , Macrolídeos/química , Macrolídeos/uso terapêutico , Esteroides/química , Esteroides/uso terapêutico , Animais , Anti-Inflamatórios/farmacocinética , Asma/induzido quimicamente , Ácidos Carboxílicos , Linhagem Celular , Desenho de Fármacos , Glucocorticoides/química , Glucocorticoides/farmacocinética , Glucocorticoides/uso terapêutico , Macrolídeos/farmacocinética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ratos , Ratos Endogâmicos BN , Esteroides/farmacocinéticaRESUMO
The murine model of cantharidin-induced ear inflammation was profiled in detail for its alignment with the human model and to explore the mechanism of anti-inflammatory activity of the macrolide antibiotics, clarithromycin and azithromycin. Ear swelling in CD1 mice persisted for 7 days, with peak intensity at 16 h after inflammation induction. As in humans, cantharidin (12.5 µg/ear) generated macrophage-inflammatory protein (MIP)-2, monocyte chemoattractant protein (MCP)-1, keratinocyte-derived chemokine (KC), interleukin (IL)-6, IL-1ß, and myeloperoxidase (MPO) production, as well as neutrophil accumulation in mouse ear tissue. The tested macrolides, clarithromycin and azithromycin, administered orally (2 × 150 mg/kg) 0.5 h before and 5 h after cantharidin challenge, reduced MIP-2, MCP-1, KC, and MPO concentrations and thereby decreased ear swelling. Our results suggest that cantharidin-induced acute inflammation represents an excellent model for translational research of novel anti-inflammatories.
Assuntos
Anti-Inflamatórios/farmacologia , Cantaridina/toxicidade , Modelos Animais de Doenças , Orelha Externa/efeitos dos fármacos , Camundongos Endogâmicos , Otite Externa , Administração Tópica , Animais , Antibacterianos/farmacologia , Azitromicina/farmacologia , Cantaridina/administração & dosagem , Claritromicina/farmacologia , Relação Dose-Resposta a Droga , Orelha Externa/imunologia , Orelha Externa/patologia , Edema/induzido quimicamente , Edema/tratamento farmacológico , Edema/imunologia , Irritantes/administração & dosagem , Irritantes/toxicidade , Masculino , Camundongos , Otite Externa/induzido quimicamente , Otite Externa/tratamento farmacológico , Otite Externa/imunologia , Pesquisa Translacional Biomédica/métodosRESUMO
A series of porphyrins, tetrapyrrole natural organic compounds, are evaluated here as endogenous anti-inflammatory agents. They directly inhibit the activity of Fyn, a non-receptor Src-family tyrosine kinase, triggering anti-inflammatory events associated with down-regulation of T-cell receptor signal transduction, leading to inhibition of tumor necrosis factor alpha (TNF-α) production. This is one of the major pro-inflammatory cytokines, associated with diseases such as diabetes, tumorigenesis, rheumatoid arthritis, and inflammatory bowel disease. Porphyrins, as a chemical class, inhibited Fyn kinase activity in a non-competitive, linear-mixed fashion. In cell-based in vitro experiments on polymorphonuclear cells, porphyrins inhibited TNF-α cytokine production, T-cell proliferation, and the generation of free radicals in the oxidative burst, in a concentration-related manner. In vivo, lipopolysaccharide-induced TNF-α production in mice was inhibited by several of the porphyrins. These findings may be very important for the overall understanding of the role(s) of porphyrins in inflammation and their possible application as new anti-inflammatory agents.
Assuntos
Anti-Inflamatórios/farmacologia , Porfirinas/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Animais , Anti-Inflamatórios/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Biologia Computacional , Humanos , Cinética , Lipopolissacarídeos/farmacologia , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Masculino , Camundongos , Simulação de Acoplamento Molecular , Porfirinas/metabolismo , Inibidores de Proteínas Quinases/metabolismo , Estrutura Terciária de Proteína , Proteínas Proto-Oncogênicas c-fyn/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-fyn/química , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Explosão Respiratória/efeitos dos fármacos , Células Sf9 , Spodoptera , Timidina/metabolismo , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Macrolide antibiotics, like erythromycin, clarithromycin, and azithromycin, possess anti-inflammatory properties. These properties are considered fundamental to the efficacy of these three macrolides in the treatment of chronic inflammatory diseases like diffuse panbronchiolitis and cystic fibrosis. However, long-term treatment with macrolide antibiotics presents a considerable risk for promotion of bacterial resistance. We have examined antibacterial and anti-inflammatory effects of a novel macrolide class: N'-substituted 2'-O,3'-N-carbonimidoyl bridged erythromycin-derived 14- and 15-membered macrolides. A small focused library was prepared, and compounds without antimicrobial activity, which inhibited IL-6 production, were selected. Data analysis led to a statistical model that could be used for the design of novel anti-inflammatory macrolides. The most promising compound from this library retained the anti-inflammatory activity observed with azithromycin in lipopolysaccharide-induced pulmonary neutrophilia in vivo. Importantly, this study strongly suggests that antimicrobial and anti-inflammatory activities of macrolides are independent and can be separated, which raises development plausibility of novel anti-inflammatory therapeutics.
Assuntos
Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Eritromicina/análogos & derivados , Interleucina-6/antagonistas & inibidores , Macrolídeos/química , Macrolídeos/farmacologia , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Anti-Inflamatórios não Esteroides/síntese química , Azitromicina/farmacologia , Azitromicina/uso terapêutico , Bronquiolite/tratamento farmacológico , Líquido da Lavagem Broncoalveolar/imunologia , Linhagem Celular , Claritromicina/farmacologia , Claritromicina/uso terapêutico , Fibrose Cística/tratamento farmacológico , Interações Medicamentosas/imunologia , Farmacorresistência Bacteriana/imunologia , Eritromicina/farmacologia , Eritromicina/uso terapêutico , Infecções por Haemophilus/tratamento farmacológico , Haemophilus influenzae/efeitos dos fármacos , Humanos , Lipopolissacarídeos/administração & dosagem , Lipopolissacarídeos/efeitos adversos , Pulmão/imunologia , Macrolídeos/síntese química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Testes de Sensibilidade Microbiana , Moraxella catarrhalis/efeitos dos fármacos , Neutrófilos/imunologia , Ratos , Ratos Sprague-Dawley , Streptococcus/efeitos dos fármacosAssuntos
Anti-Inflamatórios/farmacologia , Azitromicina/farmacologia , Endotoxemia/tratamento farmacológico , Inflamação/tratamento farmacológico , Proteína Amiloide A Sérica/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Contagem de Células Sanguíneas , Inflamação/microbiologia , Interleucina-6/sangue , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteína Amiloide A Sérica/biossínteseRESUMO
Macrolide antibiotics, including azithromycin, also possess anti-inflammatory properties. However, the molecular mechanism(s) of activity as well as the target cells for their action have not been unambiguously identified as yet. In this study, the effects of azithromycin on lipopolysaccharide (LPS)-induced pulmonary neutrophilia were investigated in mice. Using immunohistochemistry, mRNA and specific protein assays, we confirmed that azithromycin ameliorates LPS-induced pulmonary neutrophilia by inhibiting interleukin-1ß (IL-1ß) expression and production selectively in alveolar macrophages as well as in LPS-stimulated J774.2 macrophage-derived cells in vitro. Inhibition by azithromycin of neutrophilia and IL-1ß was accompanied by prevention of nuclear expression of activator protein-1 (AP-1) in both alveolar macrophages and J774.2 cells. The macrolide did not alter nuclear factor kappa B (NF-κB) or extracellular signal-regulated kinase 1/2 (ERK1/2) expression, activation or localization in LPS-stimulated lungs or in J774.2 cells. In conclusion, we have shown that inhibition of LPS-induced pulmonary neutrophilia and IL-1ß concentrations in lung tissue following azithromycin treatment is mediated through effects on alveolar macrophages. In addition, we have shown for the first time, in an in vivo model, that azithromycin inhibits AP-1 activation in alveolar macrophages, an action confirmed on J774.2 cells in vitro.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Azitromicina/farmacologia , Interleucina-1beta/biossíntese , Pulmão/imunologia , Infiltração de Neutrófilos/efeitos dos fármacos , Fator de Transcrição AP-1/antagonistas & inibidores , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , MAP Quinases Reguladas por Sinal Extracelular/imunologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Imuno-Histoquímica , Interleucina-1beta/imunologia , Lipopolissacarídeos/toxicidade , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/imunologia , NF-kappa B/metabolismo , Infiltração de Neutrófilos/imunologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Macrolide antibiotics possess immunomodulatory/anti-inflammatory properties. These properties are considered fundamental for the efficacy of macrolide antibiotics in the treatment of chronic inflammatory diseases like diffuse panbronchiolitis and cystic fibrosis. However, the molecular mechanisms and cellular targets of anti-inflammatory/immunomodulatory macrolide activity are still not fully understood. To describe anti-inflammatory effects of macrolides in more detail and to identify potential biomarkers of their activity, we have investigated the influence of azithromycin and clarithromycin on the inflammatory cascade leading to neutrophil infiltration into lungs after intranasal lipopolysaccharide challenge in mice. Azithromycin and clarithromycin pretreatment reduced total cell and neutrophil numbers in bronchoalveolar lavage fluid and myeloperoxidase concentration in lung tissue. In addition, concentrations of several inflammatory mediators, including CCL2, granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-1beta (IL-1beta), tumor necrosis factor alpha, and sE-selectin in lung homogenates were decreased after macrolide treatment. Inhibition of cytokine production observed in vivo was also corroborated in vitro in lipopolysaccharide-stimulated monocytes/macrophages, but not in an epithelial cell line. In summary, results presented in this article confirm that macrolides can suppress neutrophil-dominated pulmonary inflammation and suggest that the effect is mediated through inhibition of GM-CSF and IL-1beta production by alveolar macrophages. Besides GM-CSF and IL-1beta, CCL2 and sE-selectin are also identified as potential biomarkers of macrolide anti-inflammatory activity in the lungs.
Assuntos
Azitromicina/farmacologia , Claritromicina/farmacologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/antagonistas & inibidores , Interleucina-1beta/antagonistas & inibidores , Lipopolissacarídeos/antagonistas & inibidores , Pulmão/efeitos dos fármacos , Pulmão/patologia , Infiltração de Neutrófilos/efeitos dos fármacos , Animais , Linhagem Celular , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/fisiologia , Mediadores da Inflamação/toxicidade , Interleucina-1beta/fisiologia , Lipopolissacarídeos/toxicidade , Pulmão/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Infiltração de Neutrófilos/fisiologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/patologia , Neutrófilos/fisiologiaRESUMO
Macrolide antibiotics, a class of potent antimicrobials, also possess immunomodulatory/anti-inflammatory properties. These properties are considered fundamental for the efficacy of macrolide antibiotics in the treatment of diffuse panbronchiolitis and cystic fibrosis. In patients with asthma, macrolide antibiotics have been reported to reduce airway hyperresponsiveness and improve pulmonary function. However, their beneficial actions in asthmatics possibly could be attributed to antimicrobial activity against atypical pathogens (e.g. Chlamydia pneumoniae), corticosteroid-sparing effect (inhibition of exogenous corticosteroid metabolism), and/or their anti-inflammatory/immunomodulatory effects. In order to investigate whether efficacy of macrolide antibiotics in asthma results from their immunomodulatory/anti-inflammatory activity, the influence of clarithromycin pretreatment (2 h before challenge) was examined on ovalbumin-induced airway hyperresponsiveness and airway inflammation in the mouse. Clarithromycin treatment (200 mg/kg intraperitoneally) decreased IL-4, IL-5, IL-13, CXCL2 and CCL2 concentrations in bronchoalveolar lavage fluid and markedly reduced inflammatory cell accumulation in bronchoalveolar lavage fluid and into the lungs, as revealed by histopathological examination. Furthermore, clarithromycin-induced reduction in inflammation was accompanied by normalization of airway hyperresponsiveness. In summary, in ovalbumin-induced mouse models, clarithromycin efficiently inhibited two important pathological characteristics of asthma, airway hyperresponsiveness and inflammation. These data suggest that the efficacy of clarithromycin, as well as of other macrolide antibiotics, in asthmatic patients could be attributed to their anti-inflammatory/immunomodulatory properties, and not only to their antimicrobial activity or exogenous corticosteroid-sparing effects.
Assuntos
Asma/tratamento farmacológico , Hiper-Reatividade Brônquica/tratamento farmacológico , Claritromicina/farmacologia , Sistema Respiratório/efeitos dos fármacos , Animais , Anti-Inflamatórios/imunologia , Anti-Inflamatórios/farmacologia , Asma/imunologia , Asma/fisiopatologia , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/fisiopatologia , Líquido da Lavagem Broncoalveolar/imunologia , Claritromicina/imunologia , Modelos Animais de Doenças , Fatores Imunológicos/imunologia , Fatores Imunológicos/farmacologia , Inflamação/tratamento farmacológico , Inflamação/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Masculino , Camundongos , Ovalbumina/farmacologia , Sistema Respiratório/imunologiaRESUMO
OBJECTIVE: To investigate whether challenge with increasing allergen doses could differently affect allergen-induced airway hyperresponsiveness (AHR) and inflammatory cell accumulation in mouse model of asthma, providing an experimental model to investigate their relationship. MATERIAL AND METHODS: AHR and accumulation of inflammatory cells in bronchoalveolar lavage fluid (BALF) and into the lungs were compared in ovalbumin-sensitized mice that were challenged intranasally with 2.5, 10, 25 or 100 microg of ovalbumin/mouse. RESULTS: Both AHR and inflammatory cell accumulation were proportional to the ovalbumin dose used for challenge. However, in group challenged with 10 microg of ovalbumin airway inflammation was present, although allergen-induced AHR was not detected. Additional analysis indicated that neither mucous hyperproduction nor eosinophil degranulation could be correlated to presence of AHR in this model, whereas concentration of interleukin (IL)-13 in BALF was increased only in those groups in which AHR was present. CONCLUSIONS: Altogether, intranasal challenge of mice with increasing allergen doses could serve as a suitable experimental system for investigation of mechanisms by which airway inflammation leads to allergen-induced AHR. Our initial findings are in line with previous reports that dissociate AHR from amount of eosinophil accumulation and imply the role of IL-13 in this process.
Assuntos
Administração Intranasal , Asma/imunologia , Hiper-Reatividade Brônquica , Inflamação/imunologia , Ovalbumina , Animais , Hiper-Reatividade Brônquica/induzido quimicamente , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Modelos Animais de Doenças , Peroxidase de Eosinófilo/metabolismo , Eosinófilos/imunologia , Humanos , Interleucina-13/imunologia , Pulmão/citologia , Pulmão/imunologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/administração & dosagem , Ovalbumina/imunologiaAssuntos
Anti-Inflamatórios/farmacologia , Claritromicina , Infecções por Escherichia coli , Lipopolissacarídeos/administração & dosagem , Choque Séptico , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Animais , Anti-Inflamatórios/uso terapêutico , Claritromicina/farmacologia , Claritromicina/uso terapêutico , Modelos Animais de Doenças , Escherichia coli/metabolismo , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/mortalidade , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Choque Séptico/tratamento farmacológico , Choque Séptico/mortalidade , Resultado do TratamentoRESUMO
PURPOSE: We have assessed the use of an ultrasonic nebulization system (UNS), composed of ultrasonic nebulizer and diffusion dryer filled with charcoal, for the effective delivery of beclomethasone to the airways in a murine asthma model. METHODS: Solution of beclomethasone in ethanol was aerosolized using an ultrasonic nebulizer. Passage of the aerosol through a drying column containing charcoal and deionizer produced dry beclomethasone particles. Particles were delivered to BALB/c mice placed in a whole-body exposition chamber 1 h before intranasal challenge with ovalbumine. Efficacy of beclomethasone delivery was evaluated by examining bronchoalveolar lavage fluid (BALF) cytology. RESULTS: Effect of three UNS system parameters on aerosol particle size was investigated. The critical parameter affecting the size of dry particles was beclomethasone concentration in aerosolized solution and solution flow rate while power level of ultrasonic nebulizer generator had no effect. Administration of beclomethasone at calculated dose of 150 microg/kg to mice significantly decreased total cell number and relative eosinophil number in BALF. CONCLUSIONS: The UNS system produces a monodisperse aerosol that can be used for inhalative delivery of poorly water soluble substances to experimental animals. The UNS system minimizes formulation requirements and allows rapid and relatively simple efficacy and toxicity testing in animals.
Assuntos
Antiasmáticos/administração & dosagem , Antiasmáticos/uso terapêutico , Asma/tratamento farmacológico , Beclometasona/administração & dosagem , Beclometasona/uso terapêutico , Algoritmos , Animais , Asma/imunologia , Líquido da Lavagem Broncoalveolar/citologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nebulizadores e Vaporizadores , Ovalbumina/imunologia , Tamanho da Partícula , Pós , Hipersensibilidade Respiratória/imunologia , UltrassomRESUMO
Macrolide antibacterials inhibit the production of various cytokines and the migration of inflammatory cells. These anti-inflammatory actions of macrolides may be beneficial in attenuating inflammatory processes involved in bacterial sepsis. Therefore, we investigated the ability of azithromycin to attenuate the deleterious effects of lipopolysaccharide (LPS), in three different LPS-induced inflammatory models. Our results show that azithromycin (10 and 100 mg/kg) significantly attenuated the intraperitoneal LPS-induced increase in plasma TNF-alpha concentration. It also increased survival rate in a septic shock model in mice challenged with intravenous LPS. Oral treatment with azithromycin (up to 300 mg/kg) was less effective in suppressing neutrophil infiltration into the lungs 24 h after intranasal LPS challenge, possibly because of a slower onset of action or inadequate dosing. In the same model, azithromycin given intraperitoneally significantly improved inflammatory markers (total cell number, neutrophil percentage and MIP-2 concentration) in bronchoalveolar lavage fluid. In conclusion, azithromycin exhibits significant anti-inflammatory properties but the potency of such effects varies depending on the experimental model and route of administration.
Assuntos
Antibacterianos/farmacologia , Anti-Inflamatórios não Esteroides/farmacologia , Azitromicina/farmacologia , Lipopolissacarídeos/farmacologia , Administração Oral , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Azitromicina/administração & dosagem , Pulmão/imunologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/imunologia , Choque Séptico/metabolismo , Choque Séptico/prevenção & controle , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Interaction of iron metabolism and the immune system is complex and pathological changes in one system affect the other. Ferric sorbitol citrate (FSC), non-toxic compound of ferric ions with sorbitol and citrate. has immunomodulatory effect in treated mice. We investigated an effect of FSC on NF-kappaB expression/activation in peritoneal macrophages and spleen cells of rats. TNF-alpha concentrations in sera of control and FSC intraperitoneal (i.p.) treated Wistar rats were measured by ELISA. Furthermore, peritoneal macrophages (PM) were counted and splenocytes were isolated. PM and splenocytes were lysed and their cytoplasmic and nuclear fractions were separated by centrifugation. The influence of FSC on NF-kappaB expression and/or activity as well as expression of its inhibitor IkappaB-alpha was measured by Western blot. 1.5 and three hours after FSC treatment TNF-alpha level in sera was significantly (p < or = 0.05) increased. Activation of transcription factor NF-kappaB in PM was detected three hours after treatment, followed by significant increment in PM number. In splenocytes NF-kappaB was activated six and 48 hours after FSC application. The results indicate that, after i.p. application, FSC acts as a modulator of the immune system activating NF-kappaB in PM. PM consequently secrete TNF-alpha that activates NF-kappaB in splenocytes.
Assuntos
Citratos/farmacologia , Compostos Férricos/farmacologia , Macrófagos Peritoneais/efeitos dos fármacos , NF-kappa B/metabolismo , Baço/efeitos dos fármacos , Animais , Western Blotting , Contagem de Células , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Feminino , Proteínas I-kappa B/metabolismo , Macrófagos Peritoneais/metabolismo , Inibidor de NF-kappaB alfa , Subunidade p50 de NF-kappa B , Peritônio/citologia , Peritônio/efeitos dos fármacos , Ratos , Ratos Wistar , Baço/citologia , Baço/metabolismo , Fator de Transcrição RelA , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The effects of ferric-sorbitol-citrate and ferric-citrate on the severity of experimental arthritis, TNF-alpha secretion and the immune status were examined in mice. Arthritis was induced by footpad injection of methylated BSA and intraperitoneal injection of Bordetella pertussis. Joint and footpad swelling were measured weekly by a caliper. TNF-alpha serum levels were measured by ELISA. The immune status was determined by the response of mouse lymphocytes to ConA in vitro and by the antigen-presenting cell assay. Experimental arthritis was aggravated by ferric-citrate, whereas ferric-sorbitol-citrate did not promote it. If applied to normal (non-arthritic) mice three times a week for 4 weeks, ferric-sorbitol-citrate stimulated isolated splenocytes to increase production of TNF-alpha, the function of antigen-presenting cells and lymphocyte proliferation in response to ConA in vitro. TNF-alpha production by cultured splenocytes was also stimulated. In mice with antigen-induced arthritis, iron compounds did not additionally stimulate TNF-alpha production. Thus, we have shown that ferric-sorbitol-citrate stimulated TNF-alpha production, antigen-presenting cell activity and cellular immune response. Development of antigen-induced arthritis and TNF-alpha production in arthritic mice were not stimulated.