RESUMO
Health-care systems that develop rapidly and efficiently may increase the lifespan of humans. Nevertheless, the older population is more fragile, and is at an increased risk of disease development. A concurrently growing number of surgeries and transplantations have caused antibiotics to be used much more frequently, and for much longer periods of time, which in turn increases microbial resistance. In 1945, Fleming warned against the abuse of antibiotics in his Nobel lecture: "The time may come when penicillin can be bought by anyone in the shops. Then there is the danger that the ignorant man may easily underdose himself and by exposing his microbes to non-lethal quantities of the drug make them resistant". After 70 years, we are witnessing the fulfilment of Fleming's prophecy, as more than 700,000 people die each year due to drug-resistant diseases. Naturally occurring antimicrobial peptides protect all living matter against bacteria, and now different peptidomimetic strategies to engineer innovative antibiotics are being developed to defend humans against bacterial infections.
Assuntos
Antibacterianos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Peptídeos/uso terapêutico , Peptidomiméticos/uso terapêutico , Antibacterianos/química , Peptídeos Catiônicos Antimicrobianos/química , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Bactérias/efeitos dos fármacos , Bactérias/patogenicidade , Infecções Bacterianas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Peptídeos/químicaRESUMO
The heat shock response is a highly conserved mechanism of protection elicited in the cell by various kinds of stimuli, such as heat, sodium arsenite, oxidants and inflammation. Among the mechanisms potentially involved in mediating the protective effects of hsp, one of the most investigated is the inhibition of pro-inflammatory gene expression such as inducible nitric oxide synthase (iNOS) and inflammatory cytokines. Nevertheless, data about the effects of heat shock response on cyclooxygenase-2 expression in activated macrophages are so far not available in literature. The aim of this study was to investigate the changes in cyclooxygenase-2 expression following lipopolysaccharide stimulation of heat shocked J774 murine macrophages. We found, by Western blotting analysis and reverse transcription-polymerase chain reaction analysis (RT-PCR), that the lipopolysaccharide-induced cyclooxygenase-2 gene expression was reduced in heat shocked cells. Such a reduction was associated to activation of heat shock factor, increased levels of heat shock protein 72 and inhibition of lipopolysaccharide-induced nuclear factor-kappaB binding activity. These data suggest that the heat shock response inhibits cyclooxygenase-2 gene expression at transcriptional level, i.e. by preventing the activation of nuclear factor-kappaB, and provide additional information about mechanism(s) underlying the anti-inflammatory effect of the heat shock proteins.
Assuntos
Resposta ao Choque Térmico/fisiologia , Macrófagos/metabolismo , Prostaglandina-Endoperóxido Sintases/genética , Animais , Western Blotting , Linhagem Celular , Ciclo-Oxigenase 2 , Proteínas de Ligação a DNA/metabolismo , Dinoprostona/biossíntese , Ensaio de Desvio de Mobilidade Eletroforética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico HSP72 , Fatores de Transcrição de Choque Térmico , Proteínas de Choque Térmico/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Camundongos , NF-kappa B/metabolismo , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TranscriçãoRESUMO
The nuclear factor-kappaB (NF-kappaB) is a transcription factor that plays a pivotal role in the induction of genes involved in physiological processes, as well as in the response to inflammation. In this study, we used a selective nonpeptidyl superoxide dismutase mimetic, M40403, to investigate the role of superoxide anion in NF-kappaB activation during acute inflammation in mice. Injection of carrageenan into the pleural cavity of mice induced an acute inflammatory response characterized by fluid accumulation in the pleural cavity that contained a large number of neutrophils, as well as an increased production of tumor necrosis factor-alpha and interleukin-1beta. All parameters of inflammation were attenuated by M40403 (10 mg/kg i. p., 30 min prior to carrageenan administration). These inflammatory events were associated with the activation of NF-kappaB in the lung. In particular, the appearance of inhibitory protein kappaB-alpha (IkappaB-alpha) in homogenates of lung tissues was investigated by immunoblot analysis at 4 h after carrageenan administration. IkappaB-alpha levels were substantially reduced in the lung tissue from carrageenan-treated mice in comparison with sham-treated mice. Furthermore, to detect NF-kappaB/DNA binding activity, whole extracts from lung tissue of each mouse were analyzed by electrophoretic mobility-shift assay. The DNA binding activity significantly increased in whole extracts obtained from lung tissues of vehicle-treated mice 4 h after carrageenan administration. Treatment of mice with M40403 caused a significant inhibition of carrageenan-induced IkappaB-alpha degradation and NF-kappaB/DNA binding activity. These data confirm that M40403 exerts a potent antiinflammatory activity and clearly demonstrate that the reduction of the inflammatory process is associated with modification of the activation of signal transduction pathways.
Assuntos
Inflamação/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/fisiologia , Superóxido Dismutase/química , Superóxido Dismutase/metabolismo , Animais , Carragenina/administração & dosagem , Carragenina/imunologia , Proteínas I-kappa B/metabolismo , Interleucina-1/imunologia , Pulmão/metabolismo , Masculino , Manganês , Camundongos , Inibidor de NF-kappaB alfa , Compostos Organometálicos/metabolismo , Pleurisia/induzido quimicamente , Pleurisia/imunologia , Distribuição Aleatória , Superóxidos/metabolismo , Fator de Necrose Tumoral alfa/imunologiaRESUMO
1. The aim of this study was to evaluate whether caffeic acid phenethyl ester (CAPE), an active component of propolis, was able to reduce neointimal formation in a model of endothelial injury of rat carotid artery (balloon angioplasty). Furthermore, we investigated the relationship between neointima formation and nuclear factor-kappaB (NF-kappaB) activation and we correlated NF-kappaB activation to the expression of inducible isoform of cyclo-oxygenase (COX-2) in injured carotids. 2. In control group a significant proliferation of neointima was observed 14 days after balloon angioplasty, which was correlated to an increase of NF-kappaB/DNA binding activity as well as p50/p65 nuclear levels compared to those observed in the carotids from sham-operated rats. Furthermore, NF-kappaB activation was correlated to increased COX-2, but not beta-actin, protein expression. 3. Treatment of rats for 14 days with CAPE (3, 10, 30 mg x kg(-1)) caused a significant inhibition of all the parameters assayed, except beta-actin protein expression. 4. These results indicate that treatment with CAPE may lead to a reduction of neointima formation by inhibiting NF-kappaB activation and suggest that this agent may have therapeutic relevance for the prevention of human restenosis.
Assuntos
Ácidos Cafeicos/uso terapêutico , Lesões das Artérias Carótidas/tratamento farmacológico , Álcool Feniletílico/análogos & derivados , Álcool Feniletílico/uso terapêutico , Túnica Íntima/efeitos dos fármacos , Actinas/biossíntese , Angioplastia Coronária com Balão/efeitos adversos , Animais , Lesões das Artérias Carótidas/etiologia , Ciclo-Oxigenase 2 , Depressão Química , Ensaio de Desvio de Mobilidade Eletroforética , Isoenzimas/biossíntese , Macrófagos/efeitos dos fármacos , Macrófagos/patologia , Masculino , NF-kappa B/antagonistas & inibidores , Prostaglandina-Endoperóxido Sintases/biossíntese , Ratos , Ratos WistarRESUMO
1. The peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a member of the nuclear receptor superfamily of ligand-dependent transcription factors related to retinoid, steroid and thyroid hormone receptors. The thiazolidinedione rosiglitazone and the endogenous cyclopentenone prostaglandin (PG)D2 metabolite, 15-deoxy-Delta12,14-PGJ2 (15d-PGJ2), are two PPAR-gamma ligands, which modulate the transcription of target genes. 2. The aim of this study was to investigate the effect of rosiglitazone and 15d-PGJ2 on the tissue injury caused by ischaemia/reperfusion (I/R) of the gut. 3. I/R injury of the intestine was caused by clamping both the superior mesenteric artery and the coeliac trunk for 45 min, followed by release of the clamp allowing reperfusion for 2 or 4 h. This procedure results in splanchnic artery occlusion (SAO) shock. 4. Rats subjected to SAO developed a significant fall in mean arterial blood pressure, and only 10% of the animals survived for the entire 4 h reperfusion period. Surviving animals were killed for histological examination and biochemical studies. Rats subjected to SAO displayed a significant increase in tissue myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels, significant increases in plasma tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta levels and marked injury to the distal ileum. 5. Increased immunoreactivity to nitrotyrosine was observed in the ileum of rats subjected to SAO. Staining of sections of the ileum obtained from SAO rats with anti-intercellular adhesion molecule (ICAM-1) antibody resulted in diffuse staining. 6. Administration at 30 min prior to the onset of gut ischaemia of the two PPAR-gamma agonists (rosiglitazone (0.3 mg kg-1 i.v.) and 15d-PGJ2 (0.3 mg kg-1 i.v.)) significantly reduced the (i) fall in mean arterial blood pressure, (ii) mortality rate, (iii) infiltration of the reperfused intestine with polymorphonuclear neutrophils (MPO activity), (iv) lipid peroxidation (MDA levels), (v) production of proinflammatory cytokines (TNF-alpha and IL-1beta) and (vi) histological evidence of gut injury. Administration of rosiglitazone and 15d-PGJ2 also markedly reduced the nitrotyrosine formation and the upregulation of ICAM-1 during reperfusion. 7. In order to elucidate whether the protective effects of rosiglitazone and 15d-PGJ2 are related to the activation of the PPAR-gamma receptor, we also investigated the effect of a PPAR-gamma antagonist, bisphenol A diglycidyl ether (BADGE), on the protective effects of rosiglitazone and 15d-PGJ2. BADGE (1 mg kg-1 administered i.v. 30 min prior to the treatment of rosiglitazone or 15d-PGJ2) significantly antagonised the effect of the two PPAR-gamma agonists and thus abolished the protective effect against gut I/R. 8. These results demonstrate that the two PPAR-gamma agonists, rosiglitazone and 15d-PGJ2, significantly reduce I/R injury of the intestine.
Assuntos
Intestinos/efeitos dos fármacos , Prostaglandina D2/análogos & derivados , Prostaglandina D2/farmacologia , Receptores Citoplasmáticos e Nucleares/agonistas , Traumatismo por Reperfusão/prevenção & controle , Tiazolidinedionas/farmacologia , Fatores de Transcrição/agonistas , Tirosina/análogos & derivados , Animais , Compostos Benzidrílicos , Pressão Sanguínea/efeitos dos fármacos , Compostos de Epóxi/farmacologia , Fatores Imunológicos/farmacologia , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-1/sangue , Mucosa Intestinal/metabolismo , Intestinos/patologia , Ligantes , Masculino , Malondialdeído/metabolismo , Peroxidase/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Rosiglitazona , Fatores de Transcrição/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo , Tirosina/antagonistas & inibidores , Tirosina/biossíntese , Vasodilatadores/farmacologiaRESUMO
1. Inflammatory bowel disease (IBD) is characterized by oxidative and nitrosative stress, leukocyte infiltration, and increased expression of the adhesion molecules intercellular adhesion molecule 1 (ICAM-1) in the colon. Recent evidence also suggests that the cyclopentenone prostaglandin (PG) 15-deoxy-delta(12,14)-PGJ(2) (15d- PGJ(2)) functions as an early anti-inflammatory signal. 2. The aim of the present paper is to investigate the effects of 15d-PGJ(2) in rats subjected to experimental colitis. 3. Colitis was induced in rats by intra-colonic instillation of dinitrobenzene sulphonic acid (DNBS). 15d-PGJ(2) was administered daily as intraperitoneal injection (20 or 40 microg kg(-1)). On day 4, animals were sacrificed and tissues were taken for histological and biochemical analysis. 4. 15d-PGJ(2) significantly reduced the degree of haemorrhagic diarrhoea and weight loss caused by administration of DNBS. 15d-PGJ(2) also caused a substantial reduction of (i) the degree of colonic injury, (ii) the rise in myeloperoxidase (MPO) activity (mucosa), (iii) the increase in the tissue levels of malondialdehyde (MDA) and (iv) of the pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta). 5. Furthermore, 15d-PGJ(2) reduced the increase in immunohistochemical staining for (i) inducible nitric oxide synthase (iNOS), (ii) nitrotyrosine and (iii) poly (ADP-ribose) polymerase (PARP), as well as (iv) the increased expression of ICAM-1 caused by DNBS in the colon. 6. Electrophoresis mobility shift assay (EMSA) of inflamed colon revealed that 15d- PGJ(2) also caused a substantial reduction of the activation of nuclear factor-kappaB (NF-kappaB). Furthermore, 15d-PGJ(2) stimulates the activation of heat shock protein 72 (hsp72) in the inflamed colon, as assessed by Western blot analysis. 7. In conclusion, 15d-PGJ(2) reduces the development of experimental colitis.
Assuntos
Benzenossulfonatos/toxicidade , Colite/tratamento farmacológico , Prostaglandina D2/uso terapêutico , Animais , Colite/induzido quimicamente , Colite/metabolismo , Colite/patologia , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Masculino , Prostaglandina D2/análogos & derivados , Prostaglandina D2/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
Simplakidine A, a unique 4-alkyl-substituted pyridiunium alkaloid, has been isolated from the Caribbean sponge Plakortis simplex. The stereostructure of simplakidine A has been determined using MS and NMR data, molecular mechanics, and an extension of the J-based configuration analysis. Data about the growth-inhibition activity of simplakidine A are reported.
Assuntos
Alcaloides/química , Poríferos/química , Compostos de Piridínio/química , Alcaloides/isolamento & purificação , Alcaloides/farmacologia , Animais , Linhagem Celular , Macrófagos/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Conformação Molecular , Compostos de Piridínio/isolamento & purificação , Compostos de Piridínio/farmacologia , Espectrometria de Massas por Ionização por Electrospray/métodosRESUMO
OBJECTIVE: Nuclear factor (NF) kappaB is a transcription factor which plays a pivotal role in the induction of genes involved in physiological processes as well as in the response to injury and inflammation. Dithiocarbamates are anti-oxidants which are potent inhibitors of NF-kappaB. We postulated that pyrrolidine dithiocarbamate (PDTC) would attenuate multiple-organ failure (MOF). DESIGN AND SETTING: Rats in a university research laboratory. INTERVENTIONS AND MEASUREMENTS: We investigated the effects of PDTC (10 mg/kg) on the MOF caused by zymosan (500 mg/kg, administered i.p. as a suspension in saline) in mice. MOF in mice was assessed 18 h after administration of zymosan and/or PDTC and monitored for 7 days (for loss of body weight and mortality). RESULTS: Treatment of mice with PDTC (10 mg/kg i.p., 1 and 6 h after zymosan) attenuated the peritoneal exudation and the migration of polymorphonuclear cells caused by zymosan. PDTC also attenuated the lung, liver and pancreatic injury and renal dysfunction caused by zymosan as well as the increase in myeloperoxidase activity and malondialdehyde levels caused by zymosan in the lung, liver and intestine. Immunohistochemical analysis for inducible nitric oxide synthase, nitrotyrosine and poly(ADP-ribose) revealed positive staining in lung, liver and intestine tissues obtained from zymosan-treated mice. The degree of staining for nitrotyrosine and poly(ADP-ribose) were markedly reduced in tissue sections obtained from zymosan-treated mice which received PDTC. Furthermore, treatment of mice with PDTC significantly reduced the expression of nitric oxide synthase in lung, liver and intestine. CONCLUSIONS: This study provides the first evidence that PDTC attenuates the degree of zymosan-induced MOF in mice.
Assuntos
Antioxidantes/uso terapêutico , Modelos Animais de Doenças , Insuficiência de Múltiplos Órgãos/induzido quimicamente , Insuficiência de Múltiplos Órgãos/prevenção & controle , Pirrolidinas/uso terapêutico , Tiocarbamatos/uso terapêutico , Tirosina/análogos & derivados , Animais , Antioxidantes/farmacologia , Western Blotting , Quimiotaxia de Leucócito/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Imuno-Histoquímica , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/análise , Camundongos , Camundongos Endogâmicos , Insuficiência de Múltiplos Órgãos/diagnóstico , Insuficiência de Múltiplos Órgãos/imunologia , NF-kappa B/análise , NF-kappa B/efeitos dos fármacos , NF-kappa B/genética , NF-kappa B/imunologia , Neutrófilos/efeitos dos fármacos , Óxido Nítrico Sintase/análise , Óxido Nítrico Sintase/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II , Peroxidase/análise , Peroxidase/efeitos dos fármacos , Poli(ADP-Ribose) Polimerases/efeitos dos fármacos , Pirrolidinas/farmacologia , Distribuição Aleatória , Tiocarbamatos/farmacologia , Fatores de Tempo , Tirosina/análise , Tirosina/efeitos dos fármacos , ZimosanRESUMO
OBJECTIVE: In the present study, we investigated the effects of rosiglitazone (10 mg/kg, i.p.), a PPAR-gamma agonist, on the development of acute pancreatitis. DESIGN: Intraperitoneal injection of cerulein in mice induced an acute pancreatitis characterized by edema, neutrophil infiltration elevated serum levels of amylase and lipase. This experimental model was performed to test the anti-inflammatory activity of rosiglitazone. SETTING. University research laboratory. INTERVENTIONS: Male CD mice (20-22 g) were allocated into four groups (n=10 for each group): (a) Cerulein+vehicle group. Mice were treated hourly (x 5) with cerulein (50 microg/kg, in saline solution, i.p.); (b) Rosiglitazone group (same as the Cerulein+vehicle group but were administered rosiglitazone, 10 mg/kg bolus, 30 min prior to cerulein); (c) Sham+saline group. Mice were treated with saline instead of cerulein; (d) Sham+Rosiglitazone. Identical to Rosiglitazone group except that the saline was administered instead of cerulein. Mice were killed at 6 h after the induction of pancreatitis. Blood samples, pancreas, and lungs were collected. MEASUREMENTS AND RESULTS: Infiltration of pancreatic and lung tissue with neutrophils was associated with enhanced lipid peroxidation. Immunohistochemical examination demonstrated a marked increase in immunoreactivity for nitrotyrosine and for ICAM-1 in the pancreas of cerulein-treated mice. In contrast, the degree of (a) pancreatic inflammation and tissue injury, (b) upregulation/formation of ICAM-1 and nitrotyrosine, and (c) neutrophils infiltration was markedly reduced in pancreatic tissue obtained from rosiglitazone-treated mice. CONCLUSION: These findings support the view that rosiglitazone and other potent PPAR-gamma agonists may be useful in the therapy of acute pancreatitis.
Assuntos
Ceruletídeo/toxicidade , Pancreatite/tratamento farmacológico , Receptores Citoplasmáticos e Nucleares/agonistas , Tiazolidinedionas/uso terapêutico , Fatores de Transcrição/agonistas , Doença Aguda , Amilases/sangue , Animais , Moléculas de Adesão Celular/metabolismo , Lipase/sangue , Peroxidação de Lipídeos , Masculino , Camundongos , Pancreatite/enzimologia , Pancreatite/metabolismo , RosiglitazonaRESUMO
Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors that are related to retinoid, steroid and thyroid hormone receptors. The PPAR-gamma receptor subtype appears to play a pivotal role in the regulation of cellular proliferation and inflammation. The thiazolidinedione rosiglitazone (Avandia) is a peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist, that was recently approved by the Food and Drug Administration for treatment of type II diabetes mellitus. In the present study, we have investigated the effects of rosiglitazone in animal models of acute inflammation (carrageenan-induced paw oedema and carrageenan-induced pleurisy). We report here for the first time that rosiglitazone (given at 1, 3 or 10 mg/kg i.p. concomitantly with carrageenan injection in the paw oedema model, or at 3, 10 or 30 mg/kg i.p. 15 min before carrageenan administration in the pleurisy model) exerts potent anti-inflammatory effects (e.g. inhibition of paw oedema, pleural exudate formation, mononuclear cell infiltration and histological injury) in vivo. Furthermore, rosiglitazone reduced: (1) the increase in the staining (immunohistochemistry) for nitrotyrosine and poly (ADP-ribose) polymerase (PARP), (2) the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), intercellular adhesion molecules-1 (ICAM-1) and P-selectin in the lungs of carrageenan-treated rats. In order to elucidate whether the protective effect of rosiglitazone is related to activation of the PPAR-gamma receptor, we also investigated the effect of a PPAR-gamma antagonist, bisphenol A diglycidyl ether (BADGE), on the protective effects of rosiglitazone. BADGE (30 mg/kg i.p.) administered 30 min prior to treatment with rosiglitazone significantly antagonized the effect of the PPAR-gamma agonist and thus abolished the anti-inflammatory effects of rosiglitazone. We propose that rosiglitazone and other potent PPAR-gamma agonists may be useful in the therapy of inflammation.
Assuntos
Anti-Inflamatórios , Receptores Citoplasmáticos e Nucleares/agonistas , Tiazolidinedionas/farmacologia , Fatores de Transcrição/agonistas , Animais , Carragenina , Ciclo-Oxigenase 2 , Citocinas/biossíntese , Dinoprostona/metabolismo , Edema/induzido quimicamente , Edema/patologia , Edema/prevenção & controle , Exsudatos e Transudatos/metabolismo , Imuno-Histoquímica , Isoenzimas/biossíntese , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/patologia , Masculino , Infiltração de Neutrófilos/efeitos dos fármacos , Nitratos/metabolismo , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Nitritos/metabolismo , Peroxidase/metabolismo , Pleurisia/induzido quimicamente , Pleurisia/patologia , Pleurisia/prevenção & controle , Poli(ADP-Ribose) Polimerases/metabolismo , Prostaglandina-Endoperóxido Sintases/biossíntese , Ratos , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Rosiglitazona , Tiazolidinedionas/antagonistas & inibidores , Fatores de Transcrição/antagonistas & inibidores , Tirosina/metabolismoRESUMO
Recent studies have demonstrated that Tempol, a membrane-permeable radical scavenger, exerts protective effects in various models of inflammation and shock. Since nuclear factor-kappaB (NF-kappaB) is a transcription factor, which plays a pivotal role in the induction of genes involved in physiological processes as well as in the response to inflammation, we have investigated the effect of Tempol on NF-kappaB activation in a model of acute inflammation in mice. Injection of carrageenan into the pleural cavity of mice induced an acute inflammatory response characterized by fluid accumulation in the pleural cavity, which contained a large number of neutrophils, as well as an increased production of tumor necrosis factor-alpha, (TNF-alpha) and interleukin-1alpha (IL-1alpha). Tempol (100mg/kg i.p 30min prior to carrageenan administration) significantly attenuated the degree of pleuritis caused by carageeenan (all parameters measured). Administration of carageeenan into the chest cavity (pleuritis) was associated with the activation of NF-kappaB in the lung. In particular, the appearance of IkappaB-alpha in homogenates of lung tissue was investigated by immunoblot analysis at 4h after carrageenan administration. IkappaB-alpha levels were substantially reduced in the lung tissue from carrageenan-treated mice in comparison with sham-treated mice. Furthermore, to detect any effects of Tempol on NF-kappaB/DNA binding, lung extracts were analyzed by EMSA. The DNA binding activity significantly increased in extracts obtained from lungs of vehicle-treated mice at 4h after carrageenan administration. Treatment of mice with Tempol caused a significant inhibition of carrageenan-induced IkappaB-alpha degradation and NF-kappaB/DNA binding activity. These data confirm that Tempol exerts potent anti-inflammatory properties and clearly demonstrates for the first time that Tempol reduces the activation of NF-kappaB in vivo.
Assuntos
Óxidos N-Cíclicos/farmacologia , NF-kappa B/metabolismo , Pleurisia/metabolismo , Doença Aguda , Animais , Carragenina/farmacologia , Óxidos N-Cíclicos/uso terapêutico , DNA/metabolismo , Quinase I-kappa B , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/patologia , Interleucina-1/biossíntese , Interleucina-1/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Pleurisia/induzido quimicamente , Pleurisia/tratamento farmacológico , Pleurisia/patologia , Proteínas Serina-Treonina Quinases/metabolismo , Marcadores de Spin , Fator de Transcrição RelA , Fator de Necrose Tumoral alfa/biossíntese , Fator de Necrose Tumoral alfa/metabolismoRESUMO
In this work, the chemical compositions and antimicrobial properties of Juniperus essential oils and of their main components were determined. Five berry essential oils obtained from different species of Juniperus growing wild in Sardinia were analyzed. The components of the essential oils were identified by gas chromatography-mass spectrometry (GC-MS) analysis. The antimicrobial activities of the oils and their components against food spoilage and pathogenic microorganisms were determined by a broth microdilution method. The GC-MS analysis showed a certain variability in the concentrations of the main constituents of the oils. Alpha-pinene was largely predominant in the oils of the species J. phoenicea subsp. turbinata and J. oxycedrus. Alpha-pinene and myrcene constituted the bulk (67.56%) of the essential oil of J. communis. Significant quantitative differences were observed for myrcene, delta-3-carene, and D-germacrene. The results of the antimicrobial assay show that the oils of J. communis and J. oxycedrus failed to inhibit any of the microorganisms at the highest concentrations tested (MLC > or = 900 microg/ml), while the oils extracted from J. turbinata specimens were active against fungi, particularly against a strain of Aspergillus flavus (an aflatoxin B1 producer). Of the single compounds tested, delta-3-carene was found to possess the broadest spectrum of activity and appeared to contribute significantly to the antifungal activity observed for J. turbinata oils. This activity may be helpful in the prevention of aflatoxin contamination for many foods.
Assuntos
Antifúngicos/farmacologia , Microbiologia de Alimentos , Fungos/efeitos dos fármacos , Juniperus/química , Óleos Voláteis/farmacologia , Antifúngicos/análise , Aspergillus flavus/efeitos dos fármacos , Aspergillus flavus/crescimento & desenvolvimento , Contagem de Colônia Microbiana , Fungos/crescimento & desenvolvimento , Cromatografia Gasosa-Espectrometria de Massas , Testes de Sensibilidade Microbiana , Óleos Voláteis/análiseRESUMO
Glycoglycerolipid analogues, characterized by a glycerol aglicone beta-linked in position 2 to a glucose or galactose residue, and by a lipophilic C6 acyl chain on the glycerol unit, significantly inhibit proliferation of activated T cells. The inhibitory activity displayed by such synthetic compounds is comparable to the immunosuppressive properties shown by the natural glycolipids simplexides, to which they are structurally related. Vice versa, when the acyl chain is located on the sugar unit, no immunomodulating activity is observed, suggesting a strict relationship between the activity and the location of the acyl chain.
Assuntos
Glicolipídeos/farmacologia , Imunossupressores/farmacologia , Linfócitos T/efeitos dos fármacos , Animais , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Ativação Linfocitária , Masculino , Camundongos , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of ligand-activated transcription factors that are related to retinoid, steroid, and thyroid hormone receptors. The PPAR-gamma receptor subtype appears to play a pivotal role in the regulation of cellular proliferation and inflammation. Rosiglitazone (Avandia) is a PPAR-gamma agonist (the most potent PPAR-gamma agonist of the thiazolidinedione antidiabetics). In the present study, we investigated the effects of rosiglitazone on the development of nonseptic shock caused by zymosan in mice. DESIGN: Experimental study. SETTING: University laboratory. SUBJECTS: Male CD mice. INTERVENTIONS: We investigated the effects of rosiglitazone (3 mg/kg) on the development of nonseptic shock caused by zymosan (500 mg/kg, administered intraperitoneally as a suspension in saline) in mice. MEASUREMENTS AND MAIN RESULTS: Organ failure and systemic inflammation in rats were assessed 18 hrs after administration of zymosan and/or rosiglitazone and monitored for 12 days (for loss of body weight and mortality rate). Treatment of mice with rosiglitazone (3 mg/kg intraperitoneally, 1 and 6 hrs after zymosan) attenuated the peritoneal exudation and the migration of polymorphonuclear cells caused by zymosan. Rosiglitazone also attenuated the lung, liver, and pancreatic injury and renal dysfunction caused by zymosan as well as the increase in myeloperoxidase activity and malondialdehyde concentrations caused by zymosan in the lung and intestine. Immunohistochemical analysis for inducible nitric oxide synthase, nitrotyrosine, and poly(adenosine diphosphate-ribose) revealed positive staining in lung and intestine tissues obtained from zymosan-treated mice. The degree of staining for nitrotyrosine, inducible nitric oxide synthase, and poly(adenosine diphosphate-ribose) was markedly reduced in tissue sections obtained from zymosan-treated mice that received rosiglitazone. To elucidate whether the protective effects of rosiglitazone are related to activation of the PPAR-gamma receptor, we also investigated the effect of a PPAR-gamma antagonist, GW 9662, on the protective effects of rosiglitazone. GW 9662 (1 mg/kg administered intraperitoneally 30 mins before treatment with rosiglitazone) significantly antagonized the effect of the PPAR-gamma agonist and thus abolished the protective effect. CONCLUSIONS: This study provides evidence, for the first time, that rosiglitazone attenuates the degree of zymosan-induced nonseptic shock in mice.