IL-27 regulates IL-4-induced chemokine production in human bronchial epithelial cells.

IL-4 coordinates the Th2-type immune response in inflammatory diseases such as asthma. IL-27 can inhibit the development of both Th2 and Th1 cells. However, IL-27 can also drive naïve T cells to differentiate toward the Th1 phenotype. In this study, we investigated the effects of IL-27 on the activation of IL-4-induced human bronchial epithelial cells (BEAS-2B). Compared to controls, both IL-4 and IL-27 (25-100 ng/mL) increased the concentrations of CCL2 and IL-8 in a dose-dependent manner. However, compared to cells stimulated individually with IL-4 or IL-27, treatment with a combination of both cytokines reduced CCL2 and IL-8 concentrations in a dose- and time-dependent manner. IL-4 increased the activation of p38 MAPK, ERK1/2, STAT6 and NF-κB, while IL-27 increased the activation of p38 MAPK and ERK1/2 but not STAT6 and NF-κB. Compared to IL-4-stimulated cells, cells treated with both IL-27 and IL-4 displayed decreased activation of STAT6 and NF-κB but not ERK1/2 and p38 MAPK. Taken together, these results suggest that IL-27 plays a pro-inflammatory role when administered alone but downregulates bronchial epithelial cell activation when combined with IL-4. Therefore, IL-27 may be an interesting target for the treatment of Th2 inflammatory diseases.

Agaphelin modulates the activation of human bronchial epithelial cells induced by lipopolysaccharide and IL-4.

Sand fly saliva presents molecules with potential to development of compounds for treatment of inflammatory diseases. Agaphelin, isolated from the saliva of the mosquito Anopheles gambiae, demonstrates anti-inflammatory properties such as neutrophils chemotaxis inhibition. Here, we extend these results and evaluated the role of agaphelin (0.1-100 nM) in an in vitro model consisting in the activation of human bronchial epithelial cells (BEAS-2B) by IL-4 (50 ng/mL) or lipopolysaccharide (LPS; 10 ng/mL). Agaphelin is non-cytotoxic for BEAS-2B cells. Notably, agaphelin markedly reduces CCL2 and IL-8 production induced by IL-4 or LPS, without altering the IL-10 production. The TLR4 expression and STAT1 phosphorylation induced by LPS were inhibited by agaphlin. In addition, agaphelin decreased the phosphorylation of STAT6 induce by IL-4, whose effect was independent of IL-4-binding activity. Taken together, these findings identify agaphelin as a potential anti-inflammatory therapeutic agent for airway inflammations.

AT-RvD1 modulates the activation of bronchial epithelial cells induced by lipopolysaccharide and Dermatophagoides pteronyssinus.

Bronchial epithelial cells are essential to airways homeostasis; however, they are also involved in exacerbation of airway inflammatory responses of patients with conditions such as asthma. Dermatophagoides pteronyssinus (Dp), the most important allergen, and lipopolysaccharide (LPS), both of which are present in house dust mites (HDM), can activate immune and structural cells (such as bronchial epithelial cells) and modulate the airway inflammation in asthma patients. Resolvin D1 (RvD1) and its epimer aspirin-triggered-resolvin D1 (AT-RvD1) are lipid mediators that are produced during the resolution of inflammation and demonstrate anti-inflammatory and pro-resolution effects in several experimental models including experimental models of allergic airway inflammation. Here, we evaluated the effects of AT-RvD1 (10-12-10-10 M) on human bronchial epithelial cells (BEAS-2B) stimulated with LPS (2µg/ml) or Dp (10µg/ml). After 24h, the C-C motif chemokine ligand 2 (CCL-2) production was increased in cells that had been stimulated with LPS and Dp compared to the control. However, AT-RvD1 (10-11 and 10-10 M) significantly reduced the concentration of CCL-2 in a manner that was dependent on the N-formyl peptide receptor 2 (FPR2/ALX) and nuclear factor kappa B (NF-κB) pathways in cells stimulated with LPS or Dp compared to controls. In addition, AT-RvD1 reduced the phosphorylation of signal transducer and activator of transcription (STAT)6 and STAT1 in cells stimulated with Dp and LPS, respectively. In conclusion, AT-RvD1 demonstrated significant anti-inflammatory effects in bronchial epithelial cells that were stimulated with LPS or Dp, which provides new perspectives for therapeutic strategies to control inflammatory airway diseases.

Dieta elevada em carboidratos complexos minimiza necessidade de suplementação durante jogo-treino de rúgbi: foco no sistema imune / Diet high in complex carbohydrates minimizes necessity for supplementation during rugby training game: focus on immune system / Una dieta elevada en hidratos de carbono complejos minimiza la necesidad de suplementación durante el entrenamiento en el deporte del rugby: foco en el sistema inmunitario

Resumo O estudo analisou a imunidade oral após treino de rúgbi em atletas submetidos à dieta com alto teor de carboidratos (DATC). Em estudo randomizado, duplo-cego e placebo controlado, 20 atletas consumiram DATC por três dias antes do experimento. Os atletas receberam aleatoriamente bebida carboidratada (CHO) ou placebo (PLA) e participaram de duas sessões de treino de rúgbi, separados por sete dias. Coletas de saliva foram feitas antes (Pré-E), imediatamente após (Pós-E) e 1 h após (1 h Pós-E) o jogo-treino. Houve diferença significativa em taxa de secreção de IgA-s para PLA somente no tempo 1 h Pós-E. DATC, dias antes de treino de rúgbi, preserva função imunológica oral independente da suplementação de CHO durante treino.

Abstract The study analyzed the oral immunity after rugby training in athletes undergoing diet high in carbohydrates (DATC). In a randomized, double-blind, placebo-controlled, 20 athletes consumed DATC for three days before the experiment. The athletes were randomly carbohydrate drink (CHO) or placebo (PLA) and participated in two training sessions of rugby, separated by seven days. Saliva samples were taken before (Pre-E), immediately after (Post-E) and 1 h after (1 h post-E) training. Significant difference in rate of s-IgA secretion to PLA only at time 1 h post-E. DATC, days before training rugby, preserves immune function independent of oral CHO supplementation during training.

Resumen El estudio analizó la inmunidad oral después del entrenamiento de rugby en jugadores sometidos a una dieta elevada en hidratos de carbono (DEHC). En un estudio aleatorizado, a doble ciego, controlado con placebo, 20 jugadores consumieron una DEHC durante 3 días antes del experimento. Los jugadores recibieron aleatoriamente bebida de hidratos de carbono (CHO) o placebo (PLA), y participaron en dos sesiones de entrenamiento de rugby, separadas entre sí 7 días. Las muestras de saliva fueron antes (pre-E), inmediatamente después (post-E) y 1 hora después (1 h post-E). Hubo una diferencia significativa en la tasa de secreción de s-IgA a PLA sólo en 1 h post-E. La DEHC, días antes de la sesión de rugby, preserva la función inmunológica independientemente de la suplementación de CHO durante el entrenamiento.

Association of inflammation, dyslipidemia, obesity and physical activity status in children

The aim of this study was to verify the association between inflammatory biomarkers, dyslipidemia, obesity and physical activity status in 10-years old children. Ninety-four children participated in this study and were classified into eutrophic (n=36), overweight (n=34) or obese (n=24) according to their body mass index (BMI). The genic expression of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α) and chemokine C-C motif ligand 2 (CCL-2) mRNA; the serum concentration of high-density lipoprotein cholesterol (HDL-c) and triglycerides; BMI, percentage of body fat (% BF) and waist circumference; and the number of steps per day were determined. The expression of IL-6, TNF-α and CCL-2 were associated (p < 0.05) positively with serum triglycerides, BMI, % BF and waist circumference, and negatively with serum HDL-c. No association (p > 0.05) between pro-inflammatory biomarkers and number of steps per day was found

The anti-inflammatory and pro-resolution effects of aspirin-triggered RvD1 (AT-RvD1) on peripheral blood mononuclear cells from patients with severe asthma.

Asthma is an inflammatory disease that is characterized by a predominance of eosinophils and/or neutrophils in the airways. In the resolution of inflammation, lipid mediators such as resolvin D1 (RvD1) and its epimer aspirin-triggered RvD1 (AT-RvD1) are produced and demonstrate anti-inflammatory and pro-resolution effects. In experimental models such as airway allergic inflammation induced by ovalbumin in mice, RvD1 and AT-RvD1 alleviate some of the most important phenotypes of asthma. Here, we demonstrated the effects of AT-RvD1 on peripheral blood mononuclear cells (PBMCs) from healthy individuals and patients with severe asthma stimulated with lipopolysaccharide (LPS) or Dermatophagoides pteronyssinus (DM). AT-RvD1 (100nM) reduced the concentration of TNF-α in PBMCs from healthy individuals and patients with severe asthma stimulated with LPS or DM. In addition, AT-RvD1 lowered the production of IL-10 only in PBMCs from patients with severe asthma stimulated with LPS. These effects were associated in part with decreasing NF-κB activation. Moreover, AT-RvD1 significantly increased phagocytosis of apoptotic neutrophils by monocytes from patients with severe asthma. In conclusion, AT-RvD1 demonstrated both anti-inflammatory and pro-resolution effects in PBMCs from patients with severe asthma and could become in the future an alternative treatment for asthma.

Effects of aspirin-triggered resolvin D1 on peripheral blood mononuclear cells from patients with Chagas' heart disease.

Chagas disease is caused by Trypanosoma cruzi (T. cruzi). In some patients with Chagas disease, symptoms progress to chronic chagasic cardiomyopathy. Endogenously, inflammation is resolved in the presence of lipid mediators such as aspirin-triggered RvD1 (AT-RvD1) which has anti-inflammatory and pro-resolution effects. Here, we demonstrated, for the first time, the effects of AT-RvD1 on T. cruzi antigen-stimulated peripheral blood mononuclear cells (PBMCs) from patients with Chagas heart disease. The levels of IFN-γ, TNF-α, IL-10, and IL-13 increased in PBMCs from cardiac-form Chagas patients in stage B1 (patients with fewer heart abnormalities) stimulated with T. cruzi antigen compared to those in non-stimulated PBMCs. AT-RvD1 reduced the IFN-γ concentrations in PBMCs from patients with Chagas disease stimulated with T. cruzi antigen compared to stimulated with T. cruzi antigen cells. AT-RvD1 treatment resulted in no observable changes in TNF-α, IL-10, and IL-13 levels. AT-RvD1 significantly decreased the percentage of necrotic cells and caused a significant reduction in the proliferation rate of T. cruzi antigen-stimulated PBMCs from patients with Chagas disease. These findings demonstrate that AT-RvD1 modulates the immune response in Chagas disease patients and might have potential to be used as an alternative approach for slowing the development of further heart damage.

Anti-inflammatory Effect and Toxicology Analysis of Oral Delivery Quercetin Nanosized Emulsion in Rats.

PURPOSE: This study evaluates the advantage of the quercetin encapsulation in nanosized emulsion (QU-NE) administered orally in rats in order to demonstrate its anti-oedematous and antioxidant effects as well as its toxicity. METHODS: The nanocarriers were prepared using the hot solvent diffusion with the phase inversion temperature methods. The nanocarriers physicochemical properties were then investigated. The anti-edematous activity was tested using paw edema in rats. In addition, NF-kB expression in subcutaneous tissue of the paws was accessed by immunohistochemistry while the lipid peroxidation was analyzed in the liver by malondialdehyde reaction with thiobarbituric acid. Hematological, renal and hepatic toxicity as well as the genetic damage were also evaluated. RESULTS: The results demonstrated that QU-NE exhibited pronounced anti-oedematous property comparable to drug diclofenac. This effect was associated with NF-κB pathway inhibition. The lipid peroxidation was also only reduced in rats treated with QU-NE. Besides this, no genetic damage, hematological, renal or hepatic toxicities were observed after administration of QU-NE. CONCLUSIONS: These results suggest that quercetin nanosized emulsion exhibits anti-oedematous and antioxidant properties and does not demonstrate toxic effects. This indicates that it has a potential application in the treatment of inflammatory diseases.

AT-RvD1 modulates CCL-2 and CXCL-8 production and NF-κB, STAT-6, SOCS1, and SOCS3 expression on bronchial epithelial cells stimulated with IL-4.

Bronchial epithelial cells represent the first line of defense against microorganisms and allergens in the airways and play an important role in chronic inflammatory processes such as asthma. In an experimental model, both RvD1 and AT-RvD1, lipid mediators of inflammation resolution, ameliorated some of the most important phenotypes of experimental asthma. Here, we extend these results and demonstrate the effect of AT-RvD1 on bronchial epithelial cells (BEAS-2B) stimulated with IL-4. AT-RvD1 (100 nM) decreased both CCL2 and CXCL-8 production, in part by decreasing STAT6 and NF-κB pathways. Furthermore, the effects of AT-RvD1 were ALX/FRP2 receptor dependent, as the antagonist of this receptor (BOC1) reversed the inhibition of these chemokines by AT-RvD1. In addition, AT-RvD1 decreased SOCS1 and increased SOCS3 expression, which play important roles in Th1 and Th17 modulation, respectively. In conclusion, AT-RvD1 demonstrated significant effects on the IL-4-induced activation of bronchial epithelial cells and consequently the potential to modulate neutrophilic and eosinophilic airway inflammation in asthma. Taken together, these findings identify AT-RvD1 as a potential proresolving therapeutic agent for allergic responses in the airways.

The effects of proresolution of ellagic acid in an experimental model of allergic airway inflammation.

Asthma is a disease of airway inflammation characterized by airway hyperresponsiveness, eosinophilic inflammation, and hypersecretion of mucus. Ellagic acid, a compound derived from medicinal plants and fruits, has shown anti-inflammatory activity in several experimental disease models. We used the classical experimental model, in BALB/c mice, of sensibilization with ovalbumin to determine the effect of ellagic acid (10 mg/kg; oral route) in the resolution of allergic airways response. Dexamethasone (1 mg/kg; subcutaneous route) was used as a positive control. The control group consisted of nonimmunized mice that received challenge with ovalbumin. Ellagic acid and dexamethasone or vehicle (water) were administered before or after intranasal allergen challenge. Ellagic acid accelerated the resolution of airways inflammation by decreasing total leukocytes and eosinophils numbers in the bronchoalveolar lavage fluid (BALF), the mucus production and lung inflammation in part by reducing IL-5 concentration, eosinophil peroxidase (EPO) activity, and P-selectin expression, but not activator protein 1 (AP-1) and nuclear factor kappa B (NF-κB) pathways. In addition, ellagic acid enhanced alveolar macrophage phagocytosis of IgG-OVA-coated beads ex vivo, a new proresolving mechanism for the clearance of allergen from the airways. Together, these findings identify ellagic acid as a potential therapeutic agent for accelerating the resolution of allergic airways inflammation.

Potential effects of medicinal plants and secondary metabolites on acute lung injury.

Acute lung injury (ALI) is a life-threatening syndrome that causes high morbidity and mortality worldwide. ALI is characterized by increased permeability of the alveolar-capillary membrane, edema, uncontrolled neutrophils migration to the lung, and diffuse alveolar damage, leading to acute hypoxemic respiratory failure. Although corticosteroids remain the mainstay of ALI treatment, they cause significant side effects. Agents of natural origin, such as medicinal plants and their secondary metabolites, mainly those with very few side effects, could be excellent alternatives for ALI treatment. Several studies, including our own, have demonstrated that plant extracts and/or secondary metabolites isolated from them reduce most ALI phenotypes in experimental animal models, including neutrophil recruitment to the lung, the production of pro-inflammatory cytokines and chemokines, edema, and vascular permeability. In this review, we summarized these studies and described the anti-inflammatory activity of various plant extracts, such as Ginkgo biloba and Punica granatum, and such secondary metabolites as epigallocatechin-3-gallate and ellagic acid. In addition, we highlight the medical potential of these extracts and plant-derived compounds for treating of ALI.