Modulação por PGE Ind.3 no perfil de subpopulações celulares e citocinas na evolução do Tumor Ascítico de Ehrlich (TAE)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resposta imune induzida pelas peçonhas do bagre Cathorops agassizii

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Modulação imunológica in vitro de células de Langerhans e macrófagos por drogas utilizadas no manejo de reações hansênicas

As células de Langerhans (CLs) estão localizadas na epiderme e desempenham um papel chave na indução da resposta imune e da tolerância imunológica. Os macrófagos são células fagocíticas que atuam como primeira linha de defesa do organismo, e que estão envolvidos na formação de granulomas em pacientes com hanseníase. A imunopatogenia da resposta celular nos estados reacionais ainda é pouco estudada, porém, diversas evidências sugerem que as drogas prednisona, talidomida, ciclosporina e amitriptilina, utilizadas no controle das reações hansênicas, exercem seus efeitos pela modulação das funções de diferentes células imunocompetentes. O objetivo do presente estudo foi analisar a ação in vitro das drogas prednisona, talidomida, ciclosporina e amitriptilina sobre a produção de citocinas por CLs e macrófagos de camundongos BALB/c. As CLs foram isoladas, purificadas e cultivadas a partir da epiderme pela técnica de “panning” e os macrófagos foram isolados da cavidade peritoneal de camundongos BALB/c. Após 36 h de tratamento com as drogas, os níveis de TNF-, IL-12 e IL-10 foram medidos por ELISA. Prednisona, talidomida, ciclosporina e amitriptilina inibiram os níveis de TNF- produzidos pelas CLs, em ambas as concentrações, no entanto, não foi detectada alteração significativa na produção de IL-12. A produção de TNF- e de IL-12 por macrófagos peritoneais também foi diminuída após o tratamento, porém os níveis de IL-10 não foram modificados por nenhuma das drogas testadas. Nossos resultados mostram que estas drogas podem modular a resposta imune através da regulação das citocinas pró-inflamatórias TNF- e IL-12 por CLs purificadas da epiderme e por macrófagos peritoneais, indicando que as citocinas constituem importante alvo de drogas usadas no tratamento dos estados reacionais.

Anti-inflammatory and opioid-like activities in methanol extract of Mikania lindleyana, sucuriju

ABSTRACT: Mikania lindleyana DC., Asteraceae (sucuriju), grows in the Amazon region, where is frequently used to treat pain, inflammatory diseases and scarring. This study was carried out to investigate phytochemical profile accompanied by in vivo antinociceptive and anti-inflammatory screening of n-hexane (HE), dichloromethane (DME) and methanol (ME) extracts obtained from the aerial parts of the plant. The oral administration of ME (0.1, 0.3, 1 g/kg) caused a dose-related reduction (16.2, 42.1 e 70.2%) of acetic acid-induced abdominal writhing while HE and DME (1 g/kg, p.o.) were ineffective. In the hot plate test, ME (300 mg/kg, p.o.) increased the latency of heat stimulus between 30 and 120 min and inhibited the first (45%) and second (60%) phases of nociception in the formalin test. The antinociception induced by ME or positive control fentanyl (150 µg/kg, s.c.) in hot plate and formalin tests was prevented by naloxone (3 mg/kg, s.c.). When submitted to the carrageenan-induced peritonitis test, ME (0.5, 1.0, 2.0 g/kg, p.o.) impaired leukocyte migration into the peritoneal cavity by 46.8, 59.4 and 64.8% respectively, while positive control dexamethasone (2 mg/kg, s.c.), inhibited leukocyte migration by 71.1%. These results indicate that the antinociception obtained after oral administration of methanol extract of M. lindleyana involves anti-inflammatory mechanisms accompanied with opioid-like activity which could explain the use of the specie for pain and inflammatory diseases.

Avaliação da atividade antinociceptiva e anti-inflamatória de Mikania lindleyana DC: validação do uso na medicina popular

Mikania lindleyana (Asteraceae), popularmente conhecida como sucuriju, é uma espécie nativa da região amazônica, cujo chá das folhas é a principal forma de uso popular para tratamento de gastrite, infecção, dor e inflamação. Para validar a forma e a alegação de uso decidiu-se avaliar a toxicidade aguda e as atividades antinociceptiva e anti-inflamatória do extrato bruto aquoso liofilizado de Mikania lindleyana devidamente identificada (EAML), bem como investigar sua composição química. Na análise fitoquímica do EAML detectou-se a presença de saponinas, proteínas, aminoácidos, fenóis, taninos, ácidos orgânicos e flavonoides. Por cromatografia em camada delgada (CCD) foram observadas zonas de fluorescência azul características de ácido o-cumárico. Por cromatografia liquida acoplada à espectrometria de massa (CLAE-DAD-EM) foram encontrados compostos altamente glicosilados. O EAML na dose de 5000mg/kg não provocou morte nos animais. No teste de contorções abdominais, o EAML (nas doses 125, 250, 500, 750, 1000 e 1500mg/kg) promoveu redução no número de contorções de maneira significante e dose-dependente. A dose efetiva mediana (DE₅₀) de 692,6 mg/kg não prolongou o tempo de latência sobre a placa quente. No teste de formalina, o EAML reduziu o tempo no qual os animais permaneceram lambendo a pata injetada com formalina nas duas fases sendo este efeito revertido pelo antagonista opioide naloxona. A dose de 692,6 mg/kg inibiu a formação de eritema, mas não o edema provocado por dextrana. A mesma dose inibiu a formação do edema por carragenina a partir da 2ª hora e reduziu a migração de neutrófilos para a cavidade peritonial. Estes resultados sugerem que o EAML, nas doses utilizadas, apresenta atividade antinociceptiva na qual pode haver a participação do sistema opioide e, apresenta atividade anti-inflamatória que pode ser atribuída à ação sobre mediadores inflamatórios, como PGs, e, ainda sobre moléculas de adesão, cuja participação de citocinas pode ser crucial.

Anti-inflammatory and antinociceptive activities of Euterpe oleracea Mart., Arecaceae, oil

The oil of the fruits of Euterpe oleracea Mart., Arecaceae (OEO), was evaluated in models of inflammation and hyperalgesia in vivo to study its effects on these conditions. The experimental models contained the writhing test in mice, rat paw edema, granuloma test in rats, vascular permeability in rats, cell migration to the peritoneal cavity in rats and ear erythema induced by croton oil in mice. Doses of 500, 1000 and 1500 mg/kg of OEO were administered orally. The observed number of writhes was inhibited by 33.67, 45.88 and 55.58%, respectively. OEO produced a dose-dependent effect, with linear correlation coefficient R=0.99 (y=0.0219x+23.133), and the median effective dose found was 1226.8 mg/kg. The oral administration of 1226.8 mg/kg of OEO inhibited carrageenan-induced edema by 29.18% (p<0.05) when compared to the control group. The daily administration of OEO for six days inhibited the formation of granulomatous tissue by 36.66% (p<0.01). In ear erythema induced by croton oil, OEO presented a significant inhibition (37.9%). In the vascular permeability test, treatment with OEO decreased the response to histamine, inhibiting vascular permeability by 54.16%. In carrageenan-induced peritonitis, OEO reduced the number of neutrophils migrating compared to the control group by 80.14%. These results suggested that OEO has anti-inflammatory and antinociceptive activities, probably of peripheral origin and linked to prostaglandin biosynthesis inhibition.

Avaliação de diferentes técnicas de inseminação artificial em éguas utilizando um baixo número de espermatozóides

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sodium fluorescein for early detection of skin ulcers in Aeromonas hydrophila infected Piaractus mesopotamicus

The increase in aquaculture global fish production has been associated with an increase in infectious diseases affecting aquacultured species.. The aim of this study was to identify ulcers in pacu (Piaractus mesopotamicus) with sodium fluorescein before it was possible to observe them visually. Twenty one fish with no apparent skin lesions were divided into three groups. Group 1 were injected with saline 0.65% (Group 1), Group 2 with 6 x 10(8) CFU of Aeromonas hydrophila and group 3 with 6 x 10(8) CFU of Streptococcus agalactiae into the peritoneal cavity. All animals showed a positive reaction to fluorescein. A. hydrophila infected fish presented with large round marks with irregular borders, the characteristic ulcers caused by this agent. However, the other groups only displayed markings associated with handling caused by the netting procedures. We concluded that sodium fluorescein may be useful for an early and fast evaluation of cutaneous infection of A. hydrophila in pacu.

Estudo da migração de células peritoneais na paracoccidioidomicose experimental murina: avalaição do efeito da estimulaçao de células peritoneais por lectina sobra a evolução da lesão paracoccidióidica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diazepam, em dose única, inibe a migração celular, a estimulação macrofágica e a atividade de TNF-α na reação inflamatória aguda induzida por LPS em camundongos

Benzodiazepines are one of the most frequently prescribed drugs due to their anxiolytic properties. The aim of this study was to evaluate the effects of diazepam on lipopolysaccharide-induced peritoneal acute inflammatory responses. Swiss mice were treated with diazepam in a single dose of 1 or 10 mg/kg- subcutaneously 1 h before an intraperitoneal injection of lipopolysaccharide or sterile saline solution. The mice were killed 16 h after and the cells were washed from the peritoneal cavity to determine the total number of cells and the mononuclear and polimorfonuclear subpopulations, as well as the TNF-alpha activity and percentage of spread macrophages. Our results showed that the diazepam treatment (1 and 10 mg/kg) induced a significant reduction in the LPS-induced macrophage stimulation and TNF-α activity. Diazepam (10 mg/kg) also reduced the inflammatory cellular migration when compared to the control. It can be concluded that the diazepam treatment in a single dose is able to influence the inflammatory cellular influx, macrophage stimulation and TNF-α activity in the acute inflammatory response in mice, having possible implications on the anti-infectious response efficiency.

Modulation of cytokines production by indomethacin acute dose during the evolution of ehrlich ascites tumor in mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos da melatonina sobre a inflamação e o estresse oxidativo tecidual de pacientes submetidos à laparoscopia cirúrgica

Pós-graduação em Anestesiologia - FMB

Efeitos do pneumoperitônio e de uma manobra de recrutamento alveolar seguida por pressão positiva no final da expiração na função cardiopulmonar em ovinos anestesiados com isoflurano e fentanil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Endothelin-1 induces neutrophil recruitment in adaptive inflammation via TNF alpha and CXCL1/CXCR2 in mice

Endothelin mediates neutrophil recruitment during innate inflammation. Herein we address whether endothelin-1 (ET-1) is involved in neutrophil recruitment in adaptive inflammation in mice, and its mechanisms. Pharmacological treatments were used to determine the role of endothelin in neutrophil recruitment to the peritoneal cavity of mice challenged with antigen (ovalbumin) or ET-1. Levels of ET-1, tumour necrosis factor a (TNF alpha), and CXC chemokine ligand 1 (CXCL1) were determined by enzyme-linked immunosorbent assay. Neutrophil migration and flow cytometry analyses were performed 4 h after the intraperitoneal stimulus. ET-1 induced dose-dependent neutrophil recruitment to the peritoneal cavity. Treatment with the non-selective ETA/ETB receptor antagonist bosentan, and selective ETA or ETB receptor antagonists BQ-123 or BQ-788, respectively, inhibited ET-1- and ovalbumin-induced neutrophil migration to the peritoneal cavity. In agreement with the above, the antigen challenge significantly increased levels of ET-1 in peritoneal exudates. The ET-1- and ovalbumin-induced neutrophil recruitment were reduced in TNFR1 deficient mice, and by treatments targeting CXCL1 or CXC chemokine receptor 2 (CXCR2); further, treatment with bosentan, BQ-123, or BQ-788 inhibited ET-1- and antigen-induced production of TNFa and CXCL1. Furthermore, ET-1 and ovalbumin challenge induced an increase in the number of cells expressing the Gr1(+) markers in the granulocyte gate, CD11c+ markers in the monocyte gate, and CD4(+) and CD45(+) (B220) markers in the lymphocyte gate in an ETA-and ETB-dependent manner, as determined by flow cytometry analysis, suggesting that ET-1 might be involved in the recruitment of neutrophils and other cells in adaptive inflammation. Therefore, the present study demonstrates that ET-1 is an important mediator for neutrophil recruitment in adaptive inflammation via TNF alpha and CXCL1/CXCR2-dependent mechanism.

Hematological parameters and phagocytic activity in fat snook (Centropomus parallelus) bred in captivity

The objective of this work was to determine the hematological parameters and the phagocytic capacity of peritoneal macrophages of fat snook related to sex, stage of gonadal maturation and seasonal cycle. Blood was collected from 135 animals (78 females and 57 males) and used for determinations of: erythrocyte number, hematocrit, hemoglobin, erythrocyte indices mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), total and differential leukocyte counts, and thrombocyte count. The phagocytic capacity and phagocytic index were determined after Saccharomyces cerevisiae inoculation in the peritoneal cavity of the animals. The hematological results according to sex showed that the erythrocyte, total leukocyte and thrombocyte counts were statistically higher in males than females, with the latter showing a higher MCV. Concerning to erythrocyte count, hematocrit and hemoglobin concentration analyzed separately by sex and stage of gonadal maturation, males were found to have significantly elevated values in the mature stage and decreased levels in the resting stage. The results of the erythrocyte and leukocyte series, thrombocytes and phagocytic activity related to seasonal cycle showed significant differences in both sexes, where hematocrit and hemoglobin concentration were lower in winter and higher in the other seasons, mean corpuscular volume was higher in the summer and lower in the winter and fall, total leukocytes and thrombocytes lower in the spring and higher in the fall, lymphocytes low in the winter and summer and high in the spring and phagocytic capacity and phagocytic index high in the summer and low in the winter and fall. The results showed that the hematological values in males are statistically higher than those in females, the erythrocyte values in males increase with the progression of gonadal maturation and that winter is the season of the year least favorable for hematological and phagocytic responses for survival of fat snook kept in captivity. The parameters studied could be utilized in the evaluation of the health status of this species in captivity. (C) 2012 Elsevier Ltd. All rights reserved.