RESUMO
The objective of this study was to evaluate the development and growth of the digestive system organs, from the 11th day of incubation until the 14 d post-hatch in European and Japanese quail. On days 11, 13 and 15 of incubation at hatch and at 4, 7, 10 and 14 d post-hatch, embryos or chicks of European and Japanese quail were analyzed. After 15 d of incubation, samples from stomach and small intestine were analyzed by microscopy. European quail had significantly heavier body weight at 15 d of incubation and after 4 d post-hatch. The digestive system weight progressively increased with age and was similar between European and Japanese quail at 11, 13, and 15 d of incubation and 10 d post-hatch, while relative weight of digestive system was similar between quail type with great values at 4 d post-hatch. For relative weight of the small intestine + pancreas, the weight of the proventriculus and of the gastric ventricle increased significant by among ages analyzed in both types of quail. At hatch, proventriculus had functional secretory cells and mucosa of gastric ventricle had a thin coilin membrane. In small intestine segments, at 15 d of incubation the height of the villi was similar among duodenum, jejunum, and ileum (80 µm). Villi had elongated shape towards the intestinal lumen, covered by enterocytes and dispersed goblet cells with PAS+ and AB+ contend in all segments. The number of goblet cell/villi increased in segments until 7 to 10 d post-hatch. Duodenum increases the villi up to 14 d, while the jejunum and ileum up to 10 and 4 d, respectively. Based on our data in digestive system growth, a shorter period of post-hatch fast and specific diets to quail during first days of growth is recommended to both quail types. It is concluded that the development and growth of different organs of the digestive system up to 14 d of age was similar between European and Japanese quail.
Assuntos
Coturnix/embriologia , Coturnix/crescimento & desenvolvimento , Animais , Intestino Delgado/embriologia , Intestino Delgado/crescimento & desenvolvimento , Tamanho do Órgão , Estômago/embriologia , Estômago/crescimento & desenvolvimentoRESUMO
Fatty and hydroxycarboxylic acids are one of the main intermediates of energy metabolism in ruminants and critical in the milk production of cattle. High production demands on a dairy farm can induce nutritional imbalances and metabolism disorders, which have been widely associated with the onset of sterile inflammatory processes and increased susceptibility to infections. The literature suggests that short-chain fatty acids (SCFA), long-chain fatty acids (LCFA) and hydroxycarboxylic acids are relevant modulators of the host innate inflammatory response. For instance, increased SCFA and lactate levels are associated with subacute ruminal acidosis (SARA) and the activation of pro-inflammatory processes mediated by diverse leukocyte and vascular endothelial cells. As such, free LCFA and the ketone body ß-hydroxybutyrate are significantly increased in the plasma 1-2 weeks postpartum, coinciding with the time period in which cows are more susceptible to acquiring infectious diseases that the host innate immune system should actively oppose. Today, many of these pro-inflammatory responses can be related to the activation of specific G protein-coupled receptors, including GPR41/FFA3 and GPR43/FFA2 for SCFA; GPR40/FFA1 and GPR120/FFA4 for LCFA, GPR109A/HCA2 for ketone body ß-hydroxybutyrate, and GPR81/HCA1 for lactate, all expressed in different bovine tissues. The activation of these receptors modulates the release of intracellular granules [e.g., metalloproteinase-9 (MMP-9) and lactoferrin], radical oxygen species (ROS) production, chemotaxis, and the production of relevant pro-inflammatory mediators. The article aimed to review the role of natural ligands and receptors and the resulting impact on the host innate immune reaction of cattle and, further, to address the most recent evidence supporting a potential connection to metabolic disorders.
Assuntos
Acidose/veterinária , Ácidos Graxos não Esterificados/imunologia , Imunidade Inata , Doenças Metabólicas/veterinária , Receptores Acoplados a Proteínas G/imunologia , Acidose/metabolismo , Animais , Bovinos/imunologia , Bovinos/metabolismo , Metabolismo Energético , Ácidos Graxos Voláteis/imunologia , Feminino , Inflamação , Lactatos/metabolismo , Doenças Metabólicas/imunologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
Increased short-chain fatty acid (SCFA) production is associated with subacute ruminal acidosis (SARA) and activation of inflammatory processes. In humans and rodents, SCFAs modulate inflammatory responses in the gut via free fatty acid receptor 2 (FFA2). In bovines, butyric acid is one of the most potent FFA2 agonists. Its expression in bovine neutrophils has recently been demonstrated, suggesting a role in innate immune response in cattle. This study aimed to evaluate if butyric acid modulates oxidative and non-oxidative functions or if it can potentiate other inflammatory mediators in bovine neutrophils. Our results showed that butyric acid can activate bovine neutrophils, inducing calcium (Ca(2+)) influx and mitogen-activated protein kinase (MAPK) phosphorylation, two second messengers involved in FFA2 activation. Ca(2+) influx induced by butyric acid was dependent on the extracellular and intracellular Ca(2+) source and phospholipase C (PLC) activation. Butyric acid alone had no significant effect on reactive oxygen species (ROS) production and chemotaxis; however, a priming effect on platelet-activating factor (PAF), a potent inflammatory mediator, was observed. Butyric acid increased CD63 expression and induced the release of neutrophil granule markers matrix metalloproteinase-9 (MMP-9) and lactoferrin. Finally, we observed that butyric acid induced neutrophil extracellular trap (NET) formation without affecting cellular viability. These findings suggest that butyric acid, a component of the ruminal fermentative process, can modulate the innate immune response of ruminants.
Assuntos
Ácido Butírico/farmacologia , Bovinos/imunologia , Neutrófilos/efeitos dos fármacos , Fator de Ativação de Plaquetas/farmacologia , Animais , Cálcio/metabolismo , Quimiotaxia de Leucócito , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Armadilhas Extracelulares/efeitos dos fármacos , Ácidos Graxos não Esterificados/metabolismo , Neutrófilos/imunologia , Neutrófilos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
AIM: Neutrophils are the first cells to arrive at sites of injury. Nevertheless, many inflammatory diseases are characterized by an uncontrolled infiltration and action of these cells. Cell migration depends on volume changes that are governed by ion channel activity, but potassium channels in neutrophil have not been clearly identified. We aim to test whether KCa3.1 participates in neutrophil migration and other relevant functions of the cell. METHODS: Cytometer and confocal measurements to determine changes in cell volume were used. Cells isolated from human, mouse and horse were tested for KCa3.1-dependent chemotaxis. Chemokinetics, calcium handling and release of reactive oxygen species were measured to determine the role of KCa3.1 in those processes. A mouse model was used to test for neutrophil recruitment after acute lung injury in vivo. RESULTS: We show for the first time that KCa3.1 is expressed in mammalian neutrophils. When the channel is inhibited by a pharmacological blocker or by genetic silencing, it profoundly affects cell volume regulation, and chemotactic and chemokinetic properties of the cells. We also demonstrated that pharmacological inhibition of KCa3.1 did not affect calcium entry or reactive oxygen species production in neutrophils. Using a mouse model of acute lung injury, we observed that Kca3.1(-/-) mice are significantly less effective at recruiting neutrophils into the site of inflammation. CONCLUSIONS: These results demonstrate that KCa3.1 channels are key actors in the migration capacity of neutrophils, and its inhibition did not affect other relevant cellular functions.
Assuntos
Cálcio/metabolismo , Quimiotaxia , Canais de Potássio Ativados por Cálcio de Condutância Intermediária/metabolismo , Neutrófilos/metabolismo , Animais , Humanos , Inflamação , Potenciais da Membrana/fisiologia , Neutrófilos/citologiaRESUMO
The importance of neutrophils in human disease such as rheumatoid arthritis, asthma, adult respiratory distress syndrome, and COPD has prompted the search for drugs capable to slow down neutrophil-dependent inflammation, without interference with innate immune responses. In this review, we summarize new potential drugs targets against neutrophils mediated inflammatory responses.
Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/tratamento farmacológico , Inflamação/patologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/patologia , Animais , Humanos , Imunidade Inata/efeitos dos fármacosRESUMO
BACKGROUND: Keratinocyte life span is modulated by receptors that control proliferation and differentiation, key processes during cutaneous tissue repair. The kinin B(1) receptor (B(1)R) has been reported in normal and pathological human skin, but so far there is no information about its role in keratinocyte biology. OBJECTIVES: To determine the consequence of kinin B(1)R stimulation on tyrosine phosphorylation, a key signalling mechanism involved in keratinocyte proliferation and differentiation. METHODS: Subconfluent primary cultures of human keratinocytes were used to investigate tyrosine phosphorylation, epidermal growth factor receptor (EGFR) transactivation, cell proliferation and keratinocyte differentiation. Cell proliferation was assessed by measuring bromodeoxyuridine incorporation whereas assessment of cell differentiation was based on the expression of filaggrin, cytokeratin 10 (CK10) and involucrin. RESULTS: The major proteins phosphorylated, after B(1)R stimulation, were of molecular mass 170, 125, 89 and 70 kDa. The 170- and 125-kDa proteins were identified as EGFR and p125(FAK), respectively. Phosphorylation was greatly reduced by GF109203X and by overexposure of keratinocytes to phorbol 12-myristate 13-acetate, indicating the participation of protein kinase C. B(1)R stimulation did not increase [Ca(2+)]i, but triggered EGFR transactivation, an event that involved phosphorylation of Tyr(845), Tyr(992) and Tyr(1068) of EGFR. B(1)R stimulation did not elicit keratinocyte proliferation, but triggered cell differentiation, visualized as an increase of filaggrin, CK10 and involucrin. Blockade of EGFR tyrosine kinase by AG1478, before B(1)R stimulation, produced an additional increase in filaggrin expression. CONCLUSIONS: The kinin B(1)R may contribute to keratinocyte differentiation and migration by triggering specific tyrosine signalling pathways or by interacting with the ErbB receptor family.
Assuntos
Diferenciação Celular , Receptores ErbB/metabolismo , Queratinócitos/citologia , Cininas/metabolismo , Receptor B1 da Bradicinina/metabolismo , Receptor B2 da Bradicinina/metabolismo , Células Cultivadas/metabolismo , Proteínas Filagrinas , Humanos , Queratinócitos/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Pele/metabolismoRESUMO
Kinins are biologically active peptides that are powerful mediators of cellular inflammation. They mimic the cardinal signs of inflammation by inducing vasodilatation and by increasing vascular permeability and pain. Neutrophils are chemoattracted to sites of inflammation by several stimuli. However, the evidence concerning the chemotactic effect of kinin peptides has been contradictory. We analyzed the chemotactic effect of kinin B(1) receptor agonists on neutrophils isolated from peripheral blood of human healthy subjects. Chemotaxis was performed using the migration under agarose technique. To test the effect of B(1) receptor agonists, each assay was carried out overnight at 37 degrees C in 5% CO(2)-95% air on neutrophils primed with 1 ng/ml interleukin-1beta. Simultaneous experiments were performed using unprimed cells or cells challenged with formyl-Met-Leu-Phe (fMLP). A clear chemotactic activity was observed when primed neutrophils were challenged with Lys-des[Arg(9)]-bradykinin (LDBK) or des[Arg(9)]-bradykinin at 10(-10) M but not when unprimed cells were used. A reduction in the chemotactic response was observed after priming of cells in the presence of 0.5 mM cycloheximide and 10 mug/ml brefeldin A, suggesting that some protein biosynthesis is required. Techniques such as reverse transcriptase-polymerase chain reaction and in situ hybridization confirmed the expression of the B(1) receptor mRNA, and immunocytochemistry and autoradiography demonstrated the expression of the B(1) receptor protein. In contrast to other chemoattractants such as fMLP, cytosolic intracellular calcium did not increase in response to the B(1) receptor agonist LDBK. A generation of kinin B(1) receptor agonists during the early phase of acute inflammation may favor the recruitment of neutrophils to the inflammatory site.
Assuntos
Quimiotaxia/imunologia , Neutrófilos/imunologia , Receptor B1 da Bradicinina/imunologia , Sítios de Ligação , Antagonistas de Receptor B1 da Bradicinina , Brefeldina A/farmacologia , Diferenciação Celular/imunologia , Quimiotaxia/efeitos dos fármacos , Cicloeximida/farmacologia , Células HL-60 , Humanos , Imuno-Histoquímica , Neutrófilos/efeitos dos fármacos , Neutrófilos/ultraestrutura , RNA Mensageiro/biossíntese , RNA Mensageiro/imunologia , Receptor B1 da Bradicinina/agonistas , Relação Estrutura-AtividadeRESUMO
Injury of the renal tubulointerstitial compartment is recognized to play an important role in hypertension. Its damage may in turn, impair the activity of vasodepressor systems, like the kallikrein-kinin, in blood pressure regulation. The overload proteinuria model induces tubulointerstitial injury with activation of the renin-angiotensin system, but renal kallikrein and the development of hypertension have not received special attention. Sprague-Dawley rats received seven intraperitoneal doses of bovine serum albumin (BSA) 2 g/day under normosodic diet and were hydrated ad libitum. A second group received a high potassium diet to stimulate kallikrein production during the previous four weeks and while under BSA administration. A third one received potassium and BSA in the same schedule, but with the kinin B2 receptor antagonist, HOE140, added during the protein load phase. A control group received seven saline injections. Kallikrein protein was detected by immune labeling on renal sections and enzymatic activity in the urine. The BSA group showed massive proteinuria followed by intense tubulointerstitial damage. Blood pressure increased after the third dose in BSA animals, remaining elevated throughout the experiment, associated with significant reductions in renal expression and urinary activity of kallikrein, compared with controls. An inverse correlation was found between blood pressure and immunohistochemistry and urinary activity of kallikrein. Potassium induced a significant increase in both urinary activity and renal kallikrein expression, associated with significant reduction in blood pressure. The HOE140 antagonist blunted the antihypertensive effect of kallikrein stimulation in proteinuric rats. Loss of renal kallikrein, produced by tubulointerstitial injury, may participate in the pathogenesis of the hypertension observed in this model.
Assuntos
Calicreínas/biossíntese , Rim/metabolismo , Potássio na Dieta/administração & dosagem , Proteinúria/metabolismo , Animais , Bradicinina/análogos & derivados , Bradicinina/farmacologia , Feminino , Hipertensão/etiologia , Hipertensão/prevenção & controle , Calicreínas/urina , Rim/patologia , Proteinúria/complicações , Proteinúria/patologia , Ratos , Ratos Sprague-Dawley , Sistema Renina-Angiotensina/fisiologia , SístoleRESUMO
Tissue kallikreins are present in rat uterus during the estrous cycle in luminal and glandular epithelium, in early gestation in the implantation node, and in the last third of pregnancy surrounding the sinusoids in the decidua basalis. The pattern of kinin B2 receptor expression, through which the vasoactive effect of kallikreins is exerted, was studied by in vitro autoradiography and immunohistochemistry. The kinin B2 receptor was observed in the luminal and glandular epithelium, myometrium, endothelial cells of arteries, veins and venules, and smooth muscle cells of endometrial and myometrial arterioles. Immunoblotting of crude membranes revealed a band of 69 kDa that increased in late proestrus and estrus, concordantly with the pattern of immunostaining observed in the tissue. At Day 7 of gestation, the kinin B2 receptor was expressed (binding sites and receptor protein) in the epithelium of the implantation node and decidual cells; these latter cells showed a further increase during gestational Days 9 and 10. From Days 14 to 21, the subplacental decidua became strongly immunoreactive, and on Days 16 and 21 the placental labyrinthine endothelium was intensely stained. During this period, endothelium of arteries and veins, smooth muscular cells of small diameter arterioles, and myometrium also expressed B2 receptors. In unilaterally oil-stimulated pseudopregnancy, the decidual cells and the glandular epithelium show similar immunoreactivity to that during pregnancy. The temporospatial pattern of kinin B2 receptors, coinciding with that of kallikrein or with sites accessible to the generated kinins, further supports an autocrine-paracrine role for the kallikrein-kinin system in the vasoactive changes of implantation and placental blood flow regulation.
Assuntos
Estro/metabolismo , Prenhez/metabolismo , Receptores da Bradicinina/metabolismo , Útero/metabolismo , Animais , Autorradiografia , Bradicinina/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Endotélio Vascular/química , Epitélio/química , Feminino , Idade Gestacional , Imuno-Histoquímica , Músculo Liso Vascular , Miométrio/química , Gravidez , Pseudogravidez/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor B2 da Bradicinina , Receptores da Bradicinina/análise , Distribuição Tecidual , Útero/químicaRESUMO
Human neutrophils play a pivotal role in acute inflammation including the regulation of vascular permeability. We have examined the capacity of neutrophil enzymes to hydrolyse human kininogens in vitro and have also explored the potentiality of bradykinin to induce chemotactic migration on neutrophils isolated from peripheral blood. Isolated neutrophils were stimulated with either f-Met-Leu-Phe, thrombin or silica particles coated with human IgG. Neutrophil enzymes obtained by degranulation produced, after 45 min of incubation with high and low molecular weight kininogens, the complete transformation of both proteins in polypeptides ranging from 20 to less than 10 kDa in molecular mass. Supernatants obtained from nonstimulated neutrophils did not modify the molecular size of kininogens. The assay used to test the chemoattractant capacity of synthetic bradykinin on human neutrophils showed that this peptide has no chemotactic activity on cells isolated from healthy subjects. Our results show that stimulation of human neutrophils with opsonized silica, thrombin and the chemotactic peptide f-Met-Leu-Phe induces release of kininogen-hydrolyzing enzymes from these cells.