Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 27
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
2.
Molecules ; 27(3)2022 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-35164298

RESUMO

Certain macrolide antibiotics, azithromycin included, possess anti-inflammatory properties that are considered fundamental for their efficacy in the treatment of chronic inflammatory diseases, such as diffuse pan-bronchiolitis and cystic fibrosis. In this study, we disclose a novel azithromycin analog obtained via Barton-McCombie oxidation during which an unprecedented epimerization on the cladinose sugar occurs. Its structure was thoroughly investigated using NMR spectroscopy and compared to the natural epimer, revealing how the change in configuration of one single stereocenter (out of 16) profoundly diminished the antimicrobial activity through spatial manipulation of ribosome binding epitopes. At the same time, the anti-inflammatory properties of parent macrolide were retained, as demonstrated by inhibition of LPS- and cigarette-smoke-induced pulmonary inflammation. Not surprisingly, the compound has promising developable properties including good oral bioavailability and a half-life that supports once-daily dosing. This novel anti-inflammatory candidate has significant potential to fill the gap in existing anti-inflammatory agents and broaden treatment possibilities.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Azitromicina/análogos & derivados , Azitromicina/farmacologia , Animais , Antibacterianos/síntese química , Anti-Inflamatórios/síntese química , Azitromicina/síntese química , Bactérias/efeitos dos fármacos , Infecções Bacterianas/tratamento farmacológico , Células Cultivadas , Humanos , Macrolídeos/síntese química , Macrolídeos/química , Macrolídeos/farmacologia , Camundongos Endogâmicos BALB C , Modelos Moleculares , Oxirredução , Pneumonia/tratamento farmacológico
3.
Front Pharmacol ; 12: 682614, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34867313

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.

4.
5.
Sci Rep ; 10(1): 1104, 2020 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-31980692

RESUMO

A ten-week feeding trial was carried out to investigate the effects of replacing fishmeal (FM) with soybean meal (SBM) and brewer's yeast (BY) on growth performance, blood parameters, oxidative stress and micromorphology of liver and intestines in brown bullhead (Ameiurus nebulosus L.). Fish were fed nine feeds in which FM was replaced with 25%, 50%, 75% and 100% SBM (K1, K2, K3 and K4) and 17% + 8%, 42% + 8%, 67% + 8% and 92% + 8% of SBM/BY combination (K5, K6, K7, K8). Growth indices showed greater outcomes for the K2 group in comparison to all other groups. A decrease in plasma cholesterol and triglycerides concentrations was found after FM replacement. Activity of SOD was higher in groups K4, K7 and K8. The early inflammatory indications with abnormal vacuolization of lamina propria and basal epithelium were present in diets K4 and K8. Hepatocytes were irregular in shape with signs of inflammatory reaction in diet K8. A decreased perimeter of hepatocyte nuclei was detected in all experimental diets when compared with the control. This study demonstrates that the optimal replacement of FM with SBM/BY in brown bullhead diets contains up to 50% of FM replaced with SBM in order to obtain advantageous growth performance and adequate health condition.


Assuntos
Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Dieta/veterinária , Pesqueiros , Glycine max , Ictaluridae/crescimento & desenvolvimento , Saccharomyces cerevisiae , Animais , Colesterol/sangue , Produtos Pesqueiros , Hepatócitos/patologia , Ictaluridae/anatomia & histologia , Ictaluridae/sangue , Intestinos/anatomia & histologia , Intestinos/patologia , Fígado/anatomia & histologia , Fígado/patologia , Estresse Oxidativo , Glycine max/efeitos adversos , Superóxido Dismutase/sangue , Triglicerídeos/sangue
6.
Sci Rep ; 9(1): 5339, 2019 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-30926865

RESUMO

Staphylococcus aureus pneumonia is associated with high mortality irrespective of antibiotic susceptibility. Both MRSA and MSSA strains produce powerful cytotoxins: alpha-hemolysin(Hla) and up to five leukocidins - LukSF-PV, HlgAB, HlgCB, LukED and LukGH (LukAB) - to evade host innate defense mechanisms. Neutralizing cytotoxins has been shown to provide survival benefit in rabbit S. aureus pneumonia models. We studied the mechanisms of protection of ASN100, a combination of two human monoclonal antibodies (mAbs), ASN-1 and ASN-2, that together neutralize Hla and the five leukocidins, in rabbit MRSA and MSSA pneumonia models. Upon prophylactic passive immunization, ASN100 displayed dose-dependent increase in survival and was fully protective against all S. aureus strains tested at 5 or 20 mg/kg doses. Macroscopic and microscopic lung pathology, edema rate, and bacterial burden were evaluated 12 hours post infection and reduced by ASN100. Pharmacokinetic analysis of ASN100 in bronchoalveolar-lavage fluid from uninfected animals detected efficient penetration to lung epithelial lining fluid reaching peak levels between 24 and 48 hours post dosing that were comparable to the mAb concentration measured in serum. These data confirm that the ASN100 mAbs neutralize the powerful cytotoxins of S. aureus in the lung and prevent damage to the mucosal barrier and innate immune cells.


Assuntos
Anticorpos Neutralizantes/imunologia , Imunoglobulina G/imunologia , Imunotoxinas/imunologia , Pneumonia Estafilocócica/imunologia , Pneumonia Estafilocócica/prevenção & controle , Staphylococcus aureus/imunologia , Animais , Anticorpos Monoclonais Humanizados/administração & dosagem , Anticorpos Monoclonais Humanizados/imunologia , Biópsia , Modelos Animais de Doenças , Relação Dose-Resposta Imunológica , Imunização Passiva , Imuno-Histoquímica , Imunotoxinas/administração & dosagem , Staphylococcus aureus Resistente à Meticilina/imunologia , Pneumonia Estafilocócica/mortalidade , Pneumonia Estafilocócica/patologia , Prognóstico , Coelhos
7.
Pulm Pharmacol Ther ; 55: 75-83, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30776489

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a complex lung disease with incompletely understood pathophysiology. Effectiveness of available medicines is limited and the need for new and improved therapies remains. Due to complexity of the disease, it is difficult to develop predictable in vitro models. In this study we have described precision-cut lung slices (PCLS) prepared from bleomycin treated mice as an in vitro model for testing of novel compounds with antifibrotic activity. We have shown that PCLS during in vitro incubation retain characteristics of bleomycin model with increased expression of fibrosis related genes ACTA2 (α-smooth muscle actin), COL1A1 (collagen 1), FN1 (fibronectin 1), MMP12 (matrix metalloproteinase 12) and TIMP1 (tissue inhibitor of metalloproteinases). To further evaluate PCLS as an in vitro model, we have tested ALK5 inhibitor SB525334 which was previously shown to attenuate fibrosis in in vivo bleomycin model and nintedanib which is the FDA approved treatment for IPF. SB525334 and nintedanib inhibited expression of fibrosis related genes in PCLS from bleomycin treated mice. In addition, comparable activity profile of SB525334 was achieved in PCLS and in vivo model. Altogether these results suggest that PCLS may be a suitable in vitro model for compound testing during drug development process.


Assuntos
Modelos Animais de Doenças , Fibrose Pulmonar Idiopática/fisiopatologia , Imidazóis/farmacologia , Indóis/farmacologia , Quinoxalinas/farmacologia , Animais , Bleomicina/toxicidade , Fibrose Pulmonar Idiopática/tratamento farmacológico , Pulmão/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
8.
Drug Chem Toxicol ; 38(3): 272-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25109225

RESUMO

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 fármacos
9.
Int Immunopharmacol ; 15(3): 498-504, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23462295

RESUMO

Recent reports suggest that azithromycin can shift macrophage polarization towards the alternatively activated M2 phenotype. In order to investigate its immunomodulatory activity in vivo, the influence of azithromycin on survival and cytokine production was assessed in the LPS tolerance model which is characterized by an M2 skewed response. For induction of tolerance, mice received an intraplantar injection of 30 µg LPS, 24 h prior to intravenous challenge with 350 µg LPS. Azithromycin (100 mg/kg) was administered orally, 2 h before LPS application. Influence of treatment on survival and cytokine concentration in serum was monitored. Azithromycin alone, instead of LPS, could not induce an LPS tolerant state. However, when administered before LPS priming it significantly increased survival, which was enhanced by concomitant azithromycin before LPS challenge. Azithromycin had no effect on survival when administered only prior to the LPS challenge. Tolerance induction by LPS priming was associated, upon LPS challenge, with decreased serum concentrations of pro-inflammatory cytokines, TNFα, IL-12p40 and CCL5, and increased serum concentrations of the anti-inflammatory cytokines, IL-10 and IL-1ra. Azithromycin treatment, prior to LPS priming, further reduced serum TNFα and CCL5, yielding the greatest inhibition when the macrolide was also given prior to LPS challenge. Serum concentrations of the anti-inflammatory cytokines, IL-10 and IL-1ra, were unchanged following azithromycin treatment. In summary, we have confirmed the immunomodulatory activity of azithromycin, as reflected in its ability to augment tolerance induction to LPS, promoting increased survival and reduced pro-inflammatory cytokine production, without affecting overt inflammation to LPS or anti-inflammatory cytokine production.


Assuntos
Antibacterianos/administração & dosagem , Azitromicina/administração & dosagem , Imunomodulação , Animais , Células Cultivadas , Quimiocina CCL5/sangue , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Imunomodulação/efeitos dos fármacos , Mediadores da Inflamação/sangue , Proteína Antagonista do Receptor de Interleucina 1/sangue , Interleucina-10/sangue , Lipopolissacarídeos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator de Necrose Tumoral alfa/sangue
10.
Bioorg Med Chem ; 21(1): 321-32, 2013 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-23199485

RESUMO

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/farmacocinética
11.
Toxicol Pathol ; 40(8): 1169-87, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22659244

RESUMO

Smoking-associated chronic obstructive pulmonary disease is characterized by inflammation, changes affecting small airways, and development of emphysema. Various short- and long-term models have been introduced to investigate these processes. The aim of the present study was to identify markers of early epithelial injury/adaptation in a short-term animal model of cigarette smoke exposure. Initially, male BALB/c mice were exposed to smoke from one to five cigarettes and lung changes were assessed 4 and 24 hr after smoking cessation. Subsequently, animals were exposed to smoke from five cigarettes for 2 consecutive days and lungs investigated daily until the seventh postexposure day. Lung homogenates cytokines were determined, bronchioloalveolar fluid cells were counted, and lung tissue was analyzed by immunohistochemistry. Exposure to smoke from a single cigarette induced slight pulmonary neutrophilia. Smoke from two cigarettes additionally induced de novo expression of tight junction protein, claudin-3, by alveolar duct (AD) epithelial cells. Further increases in smoke exposure induced epithelial changes in airway progenitor regions. During the recovery period, the severity/frequency of epithelial reactions slowly decreased, coinciding with the switch from acute to a chronic inflammatory reaction. Claudin-3 and Clara cell 10 kDa protein were identified as possible markers of early tobacco smoke-induced epithelial injury along ADs.


Assuntos
Células Epiteliais Alveolares/efeitos dos fármacos , Claudina-3/metabolismo , Fumar/efeitos adversos , Uteroglobina/metabolismo , Células Epiteliais Alveolares/metabolismo , Células Epiteliais Alveolares/patologia , Animais , Biomarcadores/metabolismo , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Modelos Animais de Doenças , Exposição por Inalação , Contagem de Leucócitos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/efeitos dos fármacos , Neutrófilos/patologia , Fatores de Tempo , Nicotiana , Proteína da Zônula de Oclusão-1/metabolismo
12.
J Med Chem ; 55(13): 6111-23, 2012 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-22697905

RESUMO

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 fármacos
13.
Transl Res ; 160(2): 137-45, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22677362

RESUMO

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/métodos
14.
Eur J Pharmacol ; 691(1-3): 251-60, 2012 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-22687816

RESUMO

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/biossíntese
16.
Artigo em Inglês | MEDLINE | ID: mdl-21469013

RESUMO

The aim of this study was to investigate the mutagenic and antigenotoxic effects of different doses of the flavonoid, apigenin, alone and in combination with the antitumor drugs, cyclophosphamide and doxorubicin, in vitro and in vivo. Using bacterial reverse mutation inhibition in vitro, with and without metabolic activation, the effect of apigenin (10-400 µg/plate) was studied on genotoxicity induced by cyclophosphamide (800 µg/plate) and by doxorubicin (0.2 µg/plate). Subsequent to a dose-finding study in vivo, CD1 mice were treated with either cyclophosphamide (40 mg/kg, i.p.) or doxorubicin (5 mg/kg, i.p.) with or without co-administration of apigenin (1-100 mg/kg, p.o.). Micronuclei were determined microscopically in blood smears and glutathione peroxidase (GPX), superoxide dismutase (SOD) and total antioxidative status (TAS) in whole blood, erythrocytes and plasma, respectively. Apigenin decreased doxorubicin-induced, but not cyclophosphamide-induced mutagenicity in vitro. In vivo, apigenin caused a statistically significant decrease in micronucleus frequency in response to cyclophosphamide, possibly due to active flavonoid metabolite formation or inhibition of cyclophosphamide metabolic activation. In animals treated with apigenin and doxorubicin, a significant decrease in micronucleus frequency was not observed, probably due to interindividual variability. No changes in GPX, SOD or TAS were observed in response to either cytotoxic agents or the flavonoid, possibly due to limited metabolic transformation of the drugs at the doses used. The results of the present study provide further evidence for the chemo-preventative properties of apigenin.


Assuntos
Antineoplásicos/toxicidade , Apigenina/farmacologia , Ciclofosfamida/toxicidade , Doxorrubicina/toxicidade , Animais , Antioxidantes/química , Dano ao DNA , Glutationa Peroxidase/sangue , Camundongos , Testes de Mutagenicidade , Salmonella typhimurium , Superóxido Dismutase/sangue
17.
Int Immunopharmacol ; 11(4): 424-34, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21195124

RESUMO

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 Reversa
18.
Inflammation ; 34(5): 471-86, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20872058

RESUMO

Inflammatory and antioxidant responses, in male C57Bl6J mice, to single intranasal inoculations with live or heat-killed Streptococcus pneumoniae were studied in order to tease out differences in responses. Heat-killed bacteria elicited weak lung neutrophil infiltration and raised concentrations (peak 6-8 h), in serum or lung tissue, of CXCL1 and 2, tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), and granulocyte-macrophage-colony stimulating factor, with later increases in CCL2 and IL-1ß. Live bacteria induced profound pulmonary neutrophil infiltration and acute chemokine/cytokine elevations. After 72-96 h, live S. pneumoniae induced a delayed rise in chemokines CXCL2 and CCL2, preceded by increases in TNFα, IL-1ß, and IL-6 and mononuclear infiltration of lungs. With both live and heat-killed bacteria, alveolar epithelial type II cells and alveolar macrophages were the main sources of TNFα and IL-1ß. Only live bacteria caused an acute decrease in lung glutathione peroxidase, an increase in superoxide dismutase, and a sustained increase in serum amyloid protein A. Acute innate immune responses to live and heat-killed S. pneumoniae are similar. In response to live bacteria, inflammation is greater, accompanied by changes in antioxidant enzymes and has an additional, later mononuclear component.


Assuntos
Antioxidantes/metabolismo , Mediadores da Inflamação/metabolismo , Pneumonia Pneumocócica/metabolismo , Streptococcus pneumoniae/patogenicidade , Animais , Carga Bacteriana , Quimiocina CXCL1/metabolismo , Quimiocina CXCL2/metabolismo , Citocinas/metabolismo , Glutationa Peroxidase/metabolismo , Temperatura Alta , Imunidade Inata , Interleucina-6/metabolismo , Pulmão/imunologia , Pulmão/metabolismo , Pulmão/microbiologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pneumonia Pneumocócica/imunologia , Pneumonia Pneumocócica/microbiologia , Pneumonia Pneumocócica/patologia , Streptococcus pneumoniae/imunologia , Superóxido Dismutase/metabolismo
19.
J Pharmacol Exp Ther ; 331(1): 104-13, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19633061

RESUMO

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/fisiologia
20.
Eur J Pharmacol ; 616(1-3): 236-43, 2009 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-19560456

RESUMO

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/imunologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...