RESUMO
BACKGROUND: Inflammatory bowel diseases are characterized by chronic intestinal inflammation that leads to severe destruction of the intestinal mucosa. Therefore, the understanding of their aetiology as well as the development of new medicines is an important step for the treatment of such diseases. Consequently, the development of Lactococcus lactis strains capable of delivering a eukaryotic expression vector encoding the interleukin 4 (IL-4) of Mus musculus would represent a new strategy for the elaboration of a more effective alternative therapy against Crohn's disease. RESULTS: The murine IL-4 ORF was cloned into the eukaryotic expression vector pValac::dts. The resulting plasmid-pValac::dts::IL-4-was transfected into CHO cells so that its functionality could be evaluated in vitro. With fluorescent confocal microscopy, flow cytometry and ELISA, it was observed that pValac::dts::IL-4-transfected cells produced IL-4, while non-transfected cells and cells transfected with the empty vector did not. Then, pValac::dts::IL-4 was inserted into L. lactis MG1363 FnBPA(+) in order to evaluate the therapeutic potential of the recombinant strain against TNBS-induced colitis. Intragastric administration of L. lactis MG1363 FnBPA(+) (pValac::dts::IL-4) was able to decrease the severity of colitis, with animals showing decreased levels of IL-12, IL-6 and MPO activity; and increased levels of IL-4 and IL-10. Finally, LP-isolated cells from mice administered TNBS were immunophenotyped so that the main IL-4 and IL-10 producers were identified. Mice administered the recombinant strain presented significantly higher percentages of F4/80(+)MHCII(+)Ly6C(-)IL-4(+), F4/80(+)MHCII(+)Ly6C(-)IL-10(+), F4/80(+)MHCII(+)Ly6C(-)CD206(+)CD124(+)IL-10(+) and CD4(+)Foxp3(+)IL10(+) cells compared to the other groups. CONCLUSIONS: This study shows that L. lactis MG1363 FnBPA(+) (pValac::dts::IL-4) is a good candidate to maintain the anti-inflammatory and proinflammatory balance in the gastrointestinal tract, increasing the levels of IL-10-secreting regulatory cells and, thus, demonstrating the effectiveness of this novel DNA delivery-based strategy.
Assuntos
Vetores Genéticos , Inflamação/terapia , Doenças Inflamatórias Intestinais/terapia , Interleucina-10/metabolismo , Interleucina-4/genética , Lactococcus lactis/genética , Animais , Células CHO , Cricetulus , Citocinas/imunologia , Citocinas/metabolismo , DNA/genética , Inflamação/induzido quimicamente , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/prevenção & controle , Interleucina-4/imunologia , Interleucina-4/uso terapêutico , Mucosa Intestinal/citologia , Mucosa Intestinal/imunologia , Camundongos , Mucosa/imunologia , Mucosa/ultraestrutura , TransfecçãoRESUMO
Chronic consumption of hyperpalatable and hypercaloric foods has been pointed out as a factor associated with cognitive decline and memory impairment in obesity. In this context, the integration between peripheral and central inflammation may play a significant role in the negative effects of an obesogenic environment on memory. However, little is known about how obesity-related peripheral inflammation affects specific neurotransmission systems involved with memory regulation. Here, we test the hypothesis that chronic exposure to a highly palatable diet may cause neuroinflammation, glutamatergic dysfunction, and memory impairment. For that, we exposed C57BL/6J mice to a high sugar and butter diet (HSB) for 12 weeks, and we investigated its effects on behavior, glial reactivity, blood-brain barrier permeability, pro-inflammatory features, glutamatergic alterations, plasticity, and fractalkine-CX3CR1 axis. Our results revealed that HSB diet induced a decrease in memory reconsolidation and extinction, as well as an increase in hippocampal glutamate levels. Although our data indicated a peripheral pro-inflammatory profile, we did not observe hippocampal neuroinflammatory features. Furthermore, we also observed that the HSB diet increased hippocampal fractalkine levels, a key chemokine associated with neuroprotection and inflammatory regulation. Then, we hypothesized that the elevation on glutamate levels may saturate synaptic communication, partially limiting plasticity, whereas fractalkine levels increase as a strategy to decrease glutamatergic damage.
Assuntos
Quimiocina CX3CL1 , Hipocampo , Animais , Camundongos , Quimiocina CX3CL1/metabolismo , Dieta Hiperlipídica/efeitos adversos , Hipocampo/metabolismo , Inflamação/complicações , Camundongos Endogâmicos C57BL , Obesidade/complicações , Fármacos Atuantes sobre Aminoácidos ExcitatóriosRESUMO
Gout is a painful form of inflammatory arthritis characterized by the deposition of monosodium urate (MSU) crystals in the joints. The aim of this study was to investigate the effect of peptide P140 on the inflammatory responses in crystal-induced mouse models of gout and cell models including MSU-treated human cells. Injection of MSU crystals into the knee joint of mice induced neutrophil influx and inflammatory hypernociception. Injection of MSU crystals subcutaneously into the hind paw induced edema and increased pro-inflammatory cytokines levels. Treatment with P140 effectively reduced hypernociception, the neutrophil influx, and pro-inflammatory cytokine levels in these experimental models. Furthermore, P140 modulated neutrophils chemotaxis in vitro and increased apoptosis pathways through augmented caspase 3 activity and reduced NFκB phosphorylation. Moreover, P140 increased the production of the pro-resolving mediator annexin A1 and decreased the expression of the autophagy-related ATG5-ATG12 complex and HSPA8 chaperone protein. Overall, these findings suggest that P140 exerts a significant beneficial effect in a neutrophilic inflammation observed in the model of gout that can be of special interest in the design of new therapeutic strategies.
Assuntos
Artrite Gotosa , Gota , Camundongos , Humanos , Animais , Ácido Úrico , Fosfopeptídeos/farmacologia , Gota/tratamento farmacológico , Gota/metabolismo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Neutrófilos/metabolismo , Modelos Animais de Doenças , Artrite Gotosa/tratamento farmacológicoRESUMO
BACKGROUND: It is well known that alcohol can trigger inflammatory effects in the gastrointestinal tract (GIT), interfering with mucosal homeostasis. OBJECTIVES: This study evaluated the effectiveness of Lactococcus lactis treatment in controlling the increase in molecular biomarkers related to allergic inflammation and the effect on the diversity and abundance of the Enterobacteriaceae family in the GIT after high-dose acute administration of ethanol. METHODS: Mice received ethanol or saline solution by gavage for four consecutive days, and 24 h after the last administration, the animals were given L. lactis or M17 broth orally ad libitum for two consecutive days. The animals were subsequently sacrificed and dissected. RESULTS: L. lactis treatment was able to restore basal levels of secretory immunoglobulin A in the gastric mucosa, serum total immunoglobulin E, interleukin (IL)-4 production in gastric and intestinal tissues, and IL-10 levels in gastric tissue. L. lactis treatment encouraged the diversification of the Enterobacteriaceae population, particularly the commensal species, in the GIT. CONCLUSION: This research opens a field of studies regarding the modulatory effect of L. lactis on immunological and microbial changes induced after alcohol intake.
Assuntos
Enterobacteriaceae/metabolismo , Etanol/metabolismo , Imunoglobulina E/metabolismo , Interleucina-4/metabolismo , Lactococcus lactis/metabolismo , Administração Oral , Consumo de Bebidas Alcoólicas , Animais , Citocinas/metabolismo , Etanol/administração & dosagem , Feminino , Trato Gastrointestinal , Humanos , Imunoglobulina A/metabolismo , Imunoglobulina E/sangue , Inflamação/metabolismo , Interleucina-4/sangue , Mucosa Intestinal/metabolismo , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVES: Atherosclerosis is an underlying cause of cardiovascular disease, and obesity is one of the risk factors for atherogenesis. Although a gluten-free diet (GFD) has gained popularity as a strategy for weight loss, little is known about the effects of gluten on obesity. We have previously shown a negative effect of gluten on obesity in mice. However, its effects on atherogenesis are still unknown. Therefore, the aim of this study was to determine the effects of gluten on atherosclerosis progression during obesity. METHODS: Atherosclerosis-susceptible ApoE knockout mice were subjected to an obesogenic GFD or a diet with 4.5% gluten (GD) for 10 wk. RESULTS: Results from the study found that food intake and lipid profile were similar between the groups. However, GD promoted an increase in weight gain, adiposity, and plasma glucose. Pro-inflammatory factors such as tumor necrosis factor, interleukin-6, chemokine ligand-2, and matrix metalloproteinase-2 and -9 also were increased in the adipose tissue of gluten-fed mice. This inflammatory profile was associated with reduced phosphorylation of Akt, and consequently with the intensification of insulin resistance. The GD-enhanced vascular inflammation contributed to the worsening of atherosclerosis in the aorta and aortic root. Inflammatory cells, such as monocyte/macrophage and natural killer cells, and oxidative stress markers, such as superoxide and nitrotyrosine, were increased in atherosclerotic lesions of the GD group. Furthermore, the lesions presented higher necrotic core and lower collagen content, characterizing the less stable plaques. CONCLUSION: The gluten-containing high-fat diet was associated with a more severe proatherogenic profile than the gluten-free high-fat diet owing to increased inflammatory and oxidative status at atherosclerotic lesions in obese mice.
Assuntos
Aterosclerose , Placa Aterosclerótica , Animais , Apolipoproteínas E/genética , Aterosclerose/etiologia , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Glutens , Metaloproteinase 2 da Matriz , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Knockout para ApoE , Obesidade/etiologia , Placa Aterosclerótica/etiologiaRESUMO
It is known that high-fat diet and alcohol intake can modulate the gut microbiota and consequently affect physiological processes such as fat storage and conditional behavior. However, the effects of the interaction between high-fat diet, its withdrawal and ethanol intake in gut microbiota remain unclear. To address this question, we used an animal model in which C57BL/6 mice were fed on standard (AIN93G) or high-sugar and -butter (HSB) diet for 8 weeks. Then, a protocol of free choice between water and a 10% alcohol solution was introduced, and the HSB diet was replaced with AIN93G in two experimental groups. This model allowed us to distinguish the individual effects of HSB diet and ethanol, and the effects of its interaction on the microbiome. The interaction of those factors was the main driver in the structure changes of the fecal microbial community. HSB diet and ethanol consumption directly affected the abundance of Firmicutes and Actinobacteria phylum, and Clostridiaceae and Coriobacteriaceae family. On the other hand, we also showed that abundance of Bacteroidales_S24-7 family and the Firmicutes/Bacteroidetes ratio were affected only by HSB diet consumption and that ethanol consumption was uniquely responsible for the bacterial translocation to the liver, indicating a breaking of the gut barrier. Finally, we also pointed out that the withdrawal of the HSB diet affects the preference for alcohol and shows a structural resilience in the fecal microbiome. These results highlight the importance of the gut microbiome modulation and its possible role on the phenotype developed by animals.
Assuntos
Dieta Hiperlipídica/efeitos adversos , Etanol/farmacologia , Fezes/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Adiposidade , Animais , Bacteroidetes/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Firmicutes/efeitos dos fármacos , Preferências Alimentares/efeitos dos fármacos , Microbioma Gastrointestinal/fisiologia , Masculino , Camundongos Endogâmicos C57BLRESUMO
Obesity and ancylostomiasis are considered public health problems. Recent studies have shown that infection by intestinal helminths in obese individuals can ameliorate metabolic disorder and improve glucose tolerance by decreasing both insulin resistance and low-intensity inflammation. However, few helminth species have been studied in this context, and some modulation mechanisms still require deeper investigation. Therefore, the present work aimed to investigate the role of experimental infection with Ancylostoma ceylanicum in the modulation of the immune response in an obese experimental model. Four groups of hamsters were used as follows: two groups were submitted to a hyperlipidic and hypercaloric diet capable of inducing obesity, one infected and the other uninfected; and two normonourished control groups, one infected and one uninfected by A. ceylanicum. Biochemical, haematological, parasitological and immunological parameters were evaluated. The results demonstrated that A. ceylanicum infection accentuated weight loss in obese animals compared to normonourished animals. However, obesity reduced the recovery of worms and oviposition of the females, and both infected groups showed decreased levels of haemoglobin, albumin, iron and erythrocytes. Significant relations were observed for pathogenesis in the following cases: infection interfered in lipid metabolism, which increased levels of total cholesterol and triglycerides in the obese group, and caused a decrease in HDL levels in both groups. Obesity led to an increase in glucose levels, and the infection exacerbated this parameter in both the normonourished and obese groups. Inflammation was intensified in obese animals that showed elevated macrophage and neutrophil activation in adipose tissue, enlargement of the spleen and accumulation of lipids in the liver and faeces. Despite the decrease in IFN-γ levels, the infection did not potentiated the expression of the Foxp3, IL-10 and IL-2 transcription factor for any of the infected groups, markers that could positively compensate the host from the damage caused by obesity.
Assuntos
Ancylostoma/fisiologia , Ancilostomíase/parasitologia , Obesidade/parasitologia , Ancilostomíase/genética , Ancilostomíase/metabolismo , Animais , Colesterol/metabolismo , Cricetinae , Feminino , Glucose/metabolismo , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-12/genética , Interleucina-12/metabolismo , Fígado/metabolismo , Fígado/parasitologia , Masculino , Obesidade/genética , Obesidade/metabolismo , Oviposição , Triglicerídeos/metabolismoRESUMO
Alcoholism is a multifactorial and complex disorder responsible for 5.9% of deaths worldwide. Excessive consumption of ethanol (Et-OH) induces alcoholic liver disease (ALD), a condition comprising a spectrum of clinical signs and morphological changes, ranging from fatty liver (steatosis) to more severe forms of chronic liver injury. Secondary cofactors, such as nutritional and hepatotoxic comorbid conditions, can also contribute to liver disease development. Here we investigated the effects in the progression of ALD following short-term exposure to diet high in refined carbohydrates (HC), a high-sugar and -butter (HSB) hypercaloric diet and acute Et-OH consumption. HSB diet increased the body weight (BW) and adiposity independently of acute Et-OH consumption. HC diet did not affect BW but increased the adiposity, while acute Et-OH alone did not affect BW and adiposity. All groups of mice developed steatosis except the control group. Exposure to acute Et-OH and HSB diet increased the number of neutrophils and macrophages, and apoptosis in the liver. This combination also increased the number of circulating neutrophils and reduced mononuclear cells in the blood. Thus, short-term exposure to HSB diet and acute Et-OH intake is linked to increased liver injury. These findings offer important clues to understand the hepatic injuries associated with short exposure to hypercaloric diets and acute Et-OH.
Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Dieta Hiperlipídica/efeitos adversos , Carboidratos da Dieta/efeitos adversos , Hepatopatias Alcoólicas/patologia , Adiposidade , Alanina Transaminase/sangue , Animais , Peso Corporal , Carboidratos da Dieta/administração & dosagem , Modelos Animais de Doenças , Fígado Gorduroso/etiologia , Fígado Gorduroso/patologia , Glutationa/sangue , Fígado/efeitos dos fármacos , Fígado/patologia , Hepatopatias Alcoólicas/etiologia , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/metabolismoRESUMO
Intranasal (i.n.) administration of soluble proteins induces a state of specific unresponsiveness to subsequent immunization, known as nasal tolerance. It is thought that newborns are less susceptible to nasal tolerance induction. Recently, we have shown that feeding adult animals with a protein-free diet (Aa) resulted in their arrest at an immature immunological profile. Here, we examined the effects of the Aa diet on the development of nasal tolerance to ovalbumin (OVA) in a murine model of allergic asthma. Nasal OVA administration suppressed almost totally the OVA-induced asthma-like responses (airway eosinophilia, type 2 cytokine production, and OVA-specific IgE antibodies) in chow- or casein-fed BALB/c mice. In contrast, in Aa-fed animals the suppression of asthma-like responses by nasal OVA was partial, being effective in suppressing airway eosinophilia, but not airway type 2 cytokine or OVA-specific IgE response. We conclude that animals fed the Aa diet are more resistant to the induction of nasal tolerance. Our animal model may mimic the features of the immune system of human infants.
Assuntos
Aminoácidos/imunologia , Asma/imunologia , Proteínas Alimentares/imunologia , Tolerância Imunológica , Imunidade nas Mucosas , Mucosa Nasal/imunologia , Animais , Animais Recém-Nascidos , Modelos Animais de Doenças , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Gluten exclusion (protein complex present in many cereals) has been proposed as an option for the prevention of diseases other than coeliac disease. However, the effects of gluten-free diets on obesity and its mechanisms of action have not been studied. Thus, our objective was to assess whether gluten exclusion can prevent adipose tissue expansion and its consequences. C57BL/6 mice were fed a high-fat diet containing 4.5% gluten (Control) or no gluten (GF). Body weight and adiposity gains, leukocyte rolling and adhesion, macrophage infiltration and cytokine production in adipose tissue were assessed. Blood lipid profiles, glycaemia, insulin resistance and adipokines were measured. Expression of the PPAR-α and γ, lipoprotein lipase (LPL), hormone sensitive lipase (HSL), carnitine palmitoyl acyltransferase-1 (CPT-1), insulin receptor, GLUT-4 and adipokines were assessed in epidydimal fat. Gluten-free animals showed a reduction in body weight gain and adiposity, without changes in food intake or lipid excretion. These results were associated with up-regulation of PPAR-α, LPL, HSL and CPT-1, which are related to lipolysis and fatty acid oxidation. There was an improvement in glucose homeostasis and pro-inflammatory profile-related overexpression of PPAR-γ. Moreover, intravital microscopy showed a lower number of adhered cells in the adipose tissue microvasculature. The overexpression of PPAR-γ is related to the increase of adiponectin and GLUT-4. Our data support the beneficial effects of gluten-free diets in reducing adiposity gain, inflammation and insulin resistance. The data suggests that diet gluten exclusion should be tested as a new dietary approach to prevent the development of obesity and metabolic disorders.
Assuntos
Adiposidade/fisiologia , Dieta Livre de Glúten , Inflamação/metabolismo , Resistência à Insulina , Obesidade/metabolismo , PPAR alfa/metabolismo , PPAR gama/metabolismo , Adiponectina/metabolismo , Tecido Adiposo/metabolismo , Adiposidade/imunologia , Animais , Glicemia/metabolismo , Peso Corporal , Dieta Hiperlipídica , Inflamação/prevenção & controle , Insulina/metabolismo , Metabolismo dos Lipídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/imunologia , Regulação para CimaRESUMO
The majority of contacts with foreign antigenic materials occur on the gut mucosa, and are represented by food proteins and the autochthonous microbiota. In the present study, we replaced intact dietary proteins by equivalent amounts of amino acids from weaning on and investigated its effects on the development of the immune system of mice. Adult animals reared on a balanced protein-free diet (Aa-mice) have a poorly developed gut-associated lymphoid tissue resembling suckling mice. They also display low numbers of lamina propria cells and TCRalphabeta intraepithelial lymphocytes, and low levels of secretory IgA. Levels of circulating IgG and IgA are also reduced in Aa-mice, whereas IgM levels are normal. In vitro cytokine production by cells from several lymphoid organs shows a predominant T(h)2 profile with a high concentration of IL-10 and IL-4, and a low concentration of IFN-gamma. These parameters also resemble the immunological patterns observed in pre-weaned mice. Thus, our data clearly show that exposure to food proteins after weaning has a physiological role in the maturation of the immune system both locally and systemically.