RESUMO
The aim of this study was to evaluate the pharmacological effect of FL-6, a new immunomodulatory drug, in chronic hepatitis immunologically induced in rats via porcine-serum (PS) administration. Thirty-two male Wistar rats (150 g) were divided into 4 experimental groups: (1) Control (PBS 0.5 ml 3-times per week for 8-week); (2) FL-6 (50 ng/kg 3-times per week for 4-week); (3) Hepatitis (PS 373 mg/kg twice per week for 8-week); and (4) Hepatitis + FL-6 (doses as above). Rats were sacrificed at the end of treatment. ALT, AST, ALP and gamma-GT activities, as well as IL-6 and IL-10 levels, were evaluated in serum samples. Glutathione and malondialdehyde were also analyzed. A morphological analysis of liver tissue was carried out. The hepatitis group showed an increase in ALT (1.44-fold), AST (1.28-fold), ALP (1.83-fold), gamma-GT (3.91-fold), IL-6 (2.6-fold) and IL-10 (7.1-fold) levels when compared with controls (p < 0.05). Histopathological analysis revealed an inflammatory response characterized by inflammatory infiltrates and liver damage, which was accompanied by a reduction of 74.8% in glutathione levels (p < 0.05). However, animals with hepatitis treated with FL-6 had a reduction of ALT activity (17.74%), as well as a reduction in IL-6 (24.21%) and IL-10 (30.91%) levels (p < 0.05). These animals showed a reduction in inflammatory response characterized by a decrease in inflammatory infiltrate at the hepatic parenchyma and portal structures; livers showed less damage and a reduction of necrotic and apoptotic hepatocytes. In conclusion, the treatment with FL-6 improved liver function and reduced the inflammatory marker in rats with chronic hepatitis induced by PS-administration.
Assuntos
Hepatite Crônica/tratamento farmacológico , Fatores Imunológicos/uso terapêutico , Oligopeptídeos/uso terapêutico , Animais , Glutationa/metabolismo , Hepatite Crônica/imunologia , Hepatite Crônica/patologia , Hepatite Crônica/fisiopatologia , Fatores Imunológicos/farmacologia , Interleucina-10/sangue , Interleucina-6/sangue , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/fisiopatologia , Masculino , Oligopeptídeos/farmacologia , Ratos , Ratos WistarRESUMO
Contagious ecthyma virus (CEV) is a disease caused by a parapoxvirus, also is a potent genetic carrier with the capacity for regulating apoptosis in the cells of infected skin, a mechanism that serves for evading the immune response of the host. It has been suggested that the virus may remain in the skin and be able to cause repeated infections in the same flock. The effect of infection as well as the presence of contagious ecthyma virus was evaluated in terms of lesions and apoptosis in the skin of animals, infected both naturally and experimentally. Samples used were obtained from a naturally infected sheep, 5 goats inoculated with CEV and a negative control. Samples obtained were longitudinally sectioned and processed using photon and electron microscopy, and embedded in paraffin and araldite. Samples embedded in paraffin were sectioned in 5 microm of thickness and dyed with orange eosin-hematoxilin G and Gomori's trichrom stain, apoptosis was demonstrated by the TUNEL assay, the viral antigen was revealed using polyclonal antibodies, and the presence of lymphocytes CD4+ and CD8+, with monoclonal antibodies. The samples processed in resin were cut to obtain semi-fine sections and dyed with toluidine blue-borax, and the ultra-fine sections were impregnated with lead citrate and uranyl acetate. Observations were similar in both, the natural infected animal and the experimental group. Infiltration was observed as well as images suggestive of a process of apoptosis. The TUNEL assay demonstrated that the number of epithelial cells undergoing apoptosis diminished during the process and increased among defense cells, until they almost disappeared at the beginning of healing. Cells undergoing apoptosis were located near the sebaceous glands and pilose follicles. The infiltrated lymphocytes gradually diminished. The viral antigen was observed in cells with morphology suggestive of apoptosis, located in sebaceous glands and pilose follicles. Using electron microscopy, cells with morphology compatible with that of lymphocytes were observed to be undergoing apoptosis, but there was little evidence of viral particles.
Assuntos
Apoptose/fisiologia , Linfócitos/virologia , Vírus do Orf/fisiologia , Animais , Antígenos Virais/imunologia , Ectima Contagioso/imunologia , Ectima Contagioso/patologia , Ectima Contagioso/virologia , Cabras , Ativação Linfocitária , Linfócitos/imunologia , Linfócitos/fisiologia , Vírus do Orf/imunologia , Ovinos , Pele/citologia , Pele/patologiaRESUMO
Mycoplasma infection affects the host's immune system in different ways. In this work, a kinetic approach was used to try to determine the mechanisms by which Mycoplasma cause these effects. Experiments were performed using Balb/c mice infected with Mycoplasma pulmonis and several immunological parameters were determined. It was found that at days 10 and 15 post-infection, there were significant changes in the percentages of CD4+ and CD8 + cells, in both peripheral blood and the thymus. Significant sequential increases in concentrations of both IFN-gamma and IL-4 were detected in sera, such that at day 15, there was a peak in IFN-gamma, concentration and at day 38, IL-4 concentration also peaked. By day 46, both IFN-gamma and IL-4 fell to control levels despite continued infection. Delayed hypersensitivity (DTH) was reduced in infected animals compared to non-infected controls. A small recovery in DTH was observed at day 30, which was reduced again by day 40. Altogether, the results show features of a transitional shift from Th1 to Th2 in animals that are ultimately immunologically incompetent (in both cellular and humoral immunity). It appears to be this state of incompetence that allows the microorganism to survive and thus provides an explanation for the chronic state of the disease, which is a characteristic of Mycoplasma infection.
Assuntos
Adjuvantes Imunológicos/fisiologia , Mycoplasma/imunologia , Animais , Relação CD4-CD8 , Bovinos , Ensaio de Unidades Formadoras de Colônias , Citocinas/sangue , Hipersensibilidade Tardia/imunologia , Interferon gama/sangue , Interferon gama/imunologia , Interleucina-4/sangue , Interleucina-4/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mitógenos/farmacologia , Tamanho do Órgão/fisiologia , Fito-Hemaglutininas , Pneumonia por Mycoplasma/imunologia , Pneumonia por Mycoplasma/patologia , Soroalbumina Bovina/imunologia , Baço/citologia , Baço/imunologia , Timidina/metabolismo , Timo/citologia , Timo/imunologiaRESUMO
Malignant melanoma (MM) in children, although a rare neoplasm, can occur within a preexisting congenital melanocytic nevus (CMN). All the potential risk factors for this phenomenon are not well known, but increases in S phase and G2 + M phase of cell cycle, DNA aneuploidy, and cell cycle abnormalities in precursor lesions might be among the risk factors. Using paraffin-embedded tissue, we performed a retrospective analysis of DNA content, aneuploidy, and cell cycle by flow cytometry. Two groups of patients were analyzed: 28 children with CMN who did not developed MM, and 6 patients who further developed MM. In this second group, three patients had four biopsies done before the appearance of MM and in two patients biopsies were done after the appearance of MM. All CMN not associated with MM exhibited diploid cells only, their S phase was 11.5% (+/- 3.8), and their G2 + M phase was 2.5% (+/- 2.2). Among those patients who developed MM, 3/6 had an S phase > 15.5 and a G2 + M phase > 2.3 prior to the appearance of MM. Two out of six patients had a tetraploid DNA when MM developed and died with a disseminated MM. They had an S phase > 15.5 and their G2 + M phase was > 2.5. We propose that evaluation of DNA content and cell cycle by flow cytometry is a useful method to supplement biopsy findings in children with CMN who have lesions suspicious of developing a MM.
Assuntos
Ciclo Celular/genética , DNA de Neoplasias/análise , Melanoma/genética , Nevo Pigmentado/genética , Neoplasias Cutâneas/genética , Adolescente , Criança , Pré-Escolar , Feminino , Citometria de Fluxo/métodos , Seguimentos , Humanos , Lactente , Recém-Nascido , Masculino , Melanoma/etiologia , Melanoma/patologia , Nevo Pigmentado/complicações , Nevo Pigmentado/congênito , Nevo Pigmentado/patologia , Ploidias , Estudos Retrospectivos , Neoplasias Cutâneas/etiologia , Neoplasias Cutâneas/patologiaRESUMO
The aim of this study was to investigate the effect of acetaldehyde on the activity and expression of urokinase type plasminogen activator gene in a clone of hepatic stellate cells. CFSC-2G cells showed typical morphological changes of the stellate cell activation, which were accompanied by an increase in the amount of collagen with all doses of acetaldehyde used. The treatment of the cells with doses of 100 and 175 micromol/l acetaldehyde, produced an increase in the urokinase type plasminogen activator activity not only in the cell extract, but also in conditioned medium. However, the use of higher doses of acetaldehyde (250 and 350 micromol/l) produced an inhibitory effect on the urokinase type plasminogen activator activity. In contrast, the higher urokinase type plasminogen activator gene expression was observed with doses of 175, 250, and 350 micromol/l. Our results shown that acetaldehyde induced changes in synthesis, release, and expression of urokinase type plasminogen activator in CFSC-2G cells. Those findings suggest that the alterations in the synthesis and expression of the urokinase type plasminogen activator might be another event associated to the activation of hepatic stellate cell after exposure to hepatotoxic agents like-acetaldehyde. The role of urokinase type plasminogen activator in fibrogenesis was analyzed.