ABSTRACT
PURPOSE: To investigate the effects of neonatal malnutrition followed by nutritional replacement on the signaling mechanisms developed by the inflammasome complex by analyzing the expression of the targeted TLR2, TLR4, NLRP3, caspase-1 and release of IL-1ß and IL-18 by alveolar macrophages infected in vitro with Candida albicans. METHODS: Male Wistar rats (n = 24), 90-120 days, were suckled by mothers whose diet during lactation contained 17 % protein in the nourish group and 8 % protein in the malnourished group. After weaning, both groups were fed a normal protein diet. Macrophages were obtained after tracheostomy, through the collection of bronchoalveolar lavage fluid. The quantification of the expression levels of targets (TLR2, TLR4, NLRP3 and caspase-1) was performed by real-time RT-PCR. Production of cytokines was performed by ELISA. RESULTS: The malnourished animals during lactation showed reduced body weight from the fifth day of life, remaining until adulthood. Further, the model applied malnutrition induced a lower expression of TLR4 and caspase-1. The quantification of the TLR2 and NLRP3, as well as the release of IL-1ß and IL-18, was not different between groups of animals nourished and malnourished. The system challenged with Candida albicans showed high expression levels of all targets in the study. CONCLUSIONS: The tests demonstrate nutritional restriction during critical periods of development, although nutritional supplementation may compromise defense patterns in adulthood in a timely manner, preserving distinct signaling mechanism, so that the individual does not become widely vulnerable to infections by opportunistic pathogens.
Subject(s)
Candidiasis/metabolism , Diet, Protein-Restricted/adverse effects , Gene Expression Regulation, Developmental , Inflammasomes/metabolism , Macrophages, Alveolar/metabolism , Maternal Nutritional Physiological Phenomena , Opportunistic Infections/metabolism , Animals , Animals, Newborn , Bronchoalveolar Lavage Fluid/immunology , Bronchoalveolar Lavage Fluid/microbiology , Candida albicans/immunology , Candidiasis/immunology , Candidiasis/microbiology , Candidiasis/pathology , Caspase 1/genetics , Caspase 1/metabolism , Cells, Cultured , Down-Regulation , Female , Immunity, Innate , Inflammasomes/immunology , Lactation , Macrophage Activation/immunology , Macrophages, Alveolar/immunology , Macrophages, Alveolar/microbiology , Macrophages, Alveolar/pathology , Male , Opportunistic Infections/immunology , Opportunistic Infections/microbiology , Opportunistic Infections/pathology , Rats, Wistar , Thinness/etiology , Thinness/immunology , Thinness/microbiology , Thinness/pathology , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolismABSTRACT
OBJECTIVE: Evaluate the effects of neonatal malnutrition on the microbicidal response and viability of in vitro macrophages infected with Staphylococcus aureus sensitive/resistant to methicillin. METHODS: Male Wistar rats (n = 24) were divided into two distinct groups: nourished (rats breast-fed by mothers undergoing diet with 17% casein) and malnourished (rats breast-fed by mothers undergoing diet with 8% casein). Macrophages were recovered after surgical tracheostomy procedure by collecting bronchoalveolar lavage. Four systems were established: negative control, composed only by phagocytes; positive control, macrophages plus lipopolysaccharide; and two test systems, macrophages plus Staphylococcus aureus sensitive and resistant to methicillin. Plates were incubated at 37 °C for 24 h. After this period, tests for the analysis of cell viability and microbicidal response were performed. In the statistical analysis, the Student's t and ANOVA tests were used, accepting p < 0.05. RESULTS: The neonatal malnutrition impaired the animals' body weight. There was a lower expression of the inducible nitric oxide enzyme (iNOS), nitric oxide production, and viability of macrophages infected with methicillin-resistant Staphylococcus aureus. However, increased production of superoxide anion in the malnourished group was detected. CONCLUSION: Neonatal malnutrition focusing on critical periods of development promoted lower expression of iNOS, nitric oxide production, cell viability, and exacerbated reactive oxygen species production. The high levels of reactive oxygen species may favor the onset of serious and systemic infections with fatal outcome if associated with methicillin-resistant Staphylococcus aureus.
Subject(s)
Free Radicals/metabolism , Macrophages, Alveolar/microbiology , Malnutrition/microbiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Nitric Oxide Synthase Type II/metabolism , Staphylococcus aureus/isolation & purification , Animals , Animals, Newborn , Body Weight , Cell Survival , Diet , Lipopolysaccharides/adverse effects , Macrophages, Alveolar/cytology , Male , Malnutrition/physiopathology , Methicillin/pharmacology , Microbial Viability/drug effects , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/genetics , Phagocytes/metabolism , Rats , Rats, Wistar , Reactive Oxygen Species/metabolism , Superoxides/metabolismABSTRACT
AIM: To investigate the effects of a neonatal low-protein diet on the number of macrophages in culture and the expression/production of proteins that regulate macrophage fusion in young and adult rats. METHODS: Male Wistar rats (n = 18) were suckled by mothers fed diets containing 17 % protein (controls, C) or 8 % protein (undernourished, UN). All rats were fed a normal protein diet after weaning. Bronchoalveolar lavage was collected from 42-, 60- and 90-day-old rats. Alveolar macrophages were cultured for 4 days to assess the number of cells and the expression of cadherins, key proteins involved in macrophage fusion, by western blotting. IL-4 and IFN-γ levels in culture supernatants were measured by ELISA. RESULTS: Offspring from mothers fed a low-protein diet showed a lower body weight gain. The number of cells in cultured macrophages from UN was reduced at 42 and 60 days and increased at 90 days. IL-4 production was increased in the supernatants from UN group at 60 days but did not affect the expression of cadherins. IFN-γ production was increased in the supernatants from UN group at 42 and 60 days and reduced at 90 days. CONCLUSIONS: This study thus demonstrated that dietary restriction during lactation altered the number of alveolar macrophages in culture and the production of fusion proteins of offspring aged 42, 60 or 90 days but did not modify the expression of adhesion molecules important for the fusion of these cells.