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Lycopene is well known as an efficient natural antioxidant; however, its hydrophobic structure limits intestinal absorption through the epithelium by conventional food. Oil-based edibles containing tomato increase the bioavailability of lycopene; therefore, the preparation of emulsions emerged as a solution to prepare lycopene supplements. This work reports the preparation of a composite with lycopene and layered double hydroxides (LDH) nanoparticles. Four LDH compositions were studied of which MgAl–Cl, MgAl–CO3, ZnAl–Cl and ZnAl–CO3, were their combinations. Hydrodynamic size and zeta potential were measured in aqueous suspensions for all the composites. Lycopene maintains its antioxidant capability within the magnesium-based composites and it is synergized in the zinc-containing LDHs. The improved hydrodispersability of the composites allowed us to assess the antioxidant capacity and the oxidative stress protection in conventional aqueous assays, which are limited for traditional oily lycopene formulations. The cation composition of the LDH influenced the antioxidant capacity; however, those containing magnesium presented a non-toxic performance in the oxidative stress assays. The obtained composites demonstrated to be an alternative to design lycopene aqueous formulations or increase its bioavailability.

Mechanical ventilation (MV) is essential for the treatment of critical patients since it may provide a desired gas exchange. However, MV itself can trigger ventilator-associated lung injury in patients. We hypothesized that the mechanisms of lung injury through redox imbalance might also be associated with pulmonary inflammatory status, which has not been so far described. We tested it by delivering different tidal volumes to normal lungs undergoing MV. Healthy Wistar rats were divided into spontaneously breathing animals (control group, CG), and rats were submitted to MV (controlled ventilation mode) with tidal volumes of 4 mL/kg (MVG4), 8 mL/kg (MVG8), or 12 mL/kg (MVG12), zero end-expiratory pressure (ZEEP), and normoxia ( Fi O 2 = 21 % ) for 1 hour. After ventilation and euthanasia, arterial blood, bronchoalveolar lavage fluid (BALF), and lungs were collected for subsequent analysis. MVG12 presented lower PaCO2 and bicarbonate content in the arterial blood than CG, MVG4, and MVG8. Neutrophil influx in BALF and MPO activity in lung tissue homogenate were significantly higher in MVG12 than in CG. The levels of CCL5, TNF-α, IL-1, and IL-6 in lung tissue homogenate were higher in MVG12 than in CG and MVG4. In the lung parenchyma, the lipid peroxidation was more important in MVG12 than in CG, MVG4, and MVG8, while there was more protein oxidation in MVG12 than in CG and MVG4. The stereological analysis confirmed the histological pulmonary changes in MVG12. The association of controlled mode ventilation and high tidal volume, without PEEP and normoxia, impaired pulmonary histoarchitecture and triggered redox imbalance and lung inflammation in healthy adult rats.

Mechanical ventilation (MV) is a tool used for the treatment of patients with acute or chronic respiratory failure. However, MV is a non-physiological resource, and it can cause metabolic disorders such as release of pro-inflammatory cytokines and production of reactive oxygen species. In clinical setting, maneuvers such as sigh, are used to protect the lungs. Thus, this study aimed to evaluate the effects of sigh on oxidative stress and lung inflammation in healthy adult Wistar rats submitted to MV. Male Wistar rats were divided into four groups: control (CG), mechanical ventilation (MVG), MV set at 20 sighs/h (MVG20), and MV set at 40 sighs/h (MVG40). The MVG, MVG20, and MVG40 were submitted to MV for 1 h. After the protocol, all animals were euthanized and the blood, bronchoalveolar lavage fluid, and lungs were collected for subsequent analysis. In the arterial blood, MVG40 presented higher partial pressure of oxygen and lower partial pressure of carbon dioxide compared to control. The levels of bicarbonate in MVG20 were lower compared to CG. The neutrophil influx in bronchoalveolar lavage fluid was higher in the MVG compared to CG and MVG40. In the lung parenchyma, the lipid peroxidation was higher in MVG compared to CG, MVG20, and MVG40. Superoxide dismutase and catalase activity were higher in MVG compared to CG, MVG20, and MVG40. The levels of IL-1, IL-6, and TNF in the lung homogenate were higher in MVG compared to CG, MVG20, and MVG40. The use of sigh plays a protective role as it reduced redox imbalance and pulmonary inflammation caused by MV.

Cigarette smoke is highly toxic and is a major risk factor for airway inflammation, oxidative stress, and decline in lung function-the starting points for chronic obstructive pulmonary disease. Quercetin is a potent dietary antioxidant that displays anti-inflammatory activities. The goal of this study was to evaluate the effects of quercetin on reducing the redox imbalance and inflammation induced by short-term cigarette smoke exposure.

Mechanical ventilation (MV) is a tool used in critical patient care. However, it can trigger inflammatory and oxidative processes capable of causing or aggravating lung injuries, which is known as ventilator-induced lung injury (VILI). Hesperidin is a flavonoid with antioxidant and anti-inflammatory properties in various diseases. The role of hesperidin in the process triggered by MV is poorly studied. Thus, we hypothesize hesperidin could protect the lung of mice submitted to mechanical ventilation. For that, we evaluated cell viability and reactive oxygen species (ROS) formation in macrophages using different hesperidin concentrations. We observed hesperidin did not reduce cell viability, however; it attenuated the production of intracellular ROS in cells stimulated with lipopolysaccharide (LPS). We further evaluated the effects of hesperidin in vivo in animals submitted to MV. In the bronchoalveolar lavage fluid, there were higher levels of macrophage, lymphocyte and neutrophil counts in animals submitted to MV, indicating an inflammatory process. In the lung tissue, MV induced oxidative damage and increased myeloperoxidase activity, though the antioxidant enzyme activity decreased. MV also induced the production of the inflammatory mediators CCL-2, TNF-α and IL-12. Pretreatment with hesperidin resulted in less recruitment of inflammatory cells to the airways and less oxidative damage. Also, it reduced the formation of CCL-2 and IL-12. Our results show pretreatment with hesperidin can protect the lungs of mice submitted to mechanical ventilation by modulating the inflammatory response and redox imbalance and may act to prevent MV injury.

Cigarette smoking throughout life causes serious health issues in the lungs. The electronic cigarette (E-Cig) use increased, since it was first introduced in the world. This research work compared the short-term exposure consequences to e-cigarette vapor and cigarette smoke in male mice. Forty-five C57BL/6 mice were randomized into control (C) in an ambient air exposition cigarette smoke (CS) and aerosol electronic cigarette (EC), both were exposed to 120 puffs, 3 times/day during five days. Then, in the experimental protocol, the euthanized mice had their tissues removed for analysis. Our study showed that CS and EC resulted in higher cell influx into the airways, and an increase in macrophage counts in CS (209.25 ± 7.41) and EC (220.32 ± 8.15) when compared to C (108.40 ± 4.49) ( p < 0.0001 ). The CS (1.92 ± 0.23) displayed a higher pulmonary lipid peroxidation as opposed to C (0.93 ± 0.06) and EC (1.23 ± 0.17) ( p < 0.05 ). The EC (282.30 ± 25.68) and CS (368.50 ± 38.05) promoted increased levels of interleukin 17 when compared to C (177.20 ± 10.49) ( p < 0.05 ). The EC developed shifts in lung histoarchitecture, characterized by a higher volume density in the alveolar air space (60.21; 55.00-65.83) related to C (51.25; 18.75-68.75) and CS (50.26; 43.75-62.08) (p =0.002). The EC (185.6 ± 9.01) presented a higher respiratory rate related to CS (133.6 ± 10.2) ( p < 0.002 ). Therefore, our findings demonstrated that the short-term exposure to e-cig promoted more acute inflammation comparing to cigarette smoke in the ventilatory parameters of the animals.

Introduction: Pulmonary function testing, or spirometry, is a validated, globally recognized test that contributes to the diagnosis, staging, and longitudinal follow-up of lung diseases. The exam is most often performed in a sitting position in clinical practice; hence, there are no predicted values for its performance in other positions, such as in different decubitus. Objective: The present study aimed to evaluate the effects of position on pulmonary function test results in healthy adults. Methods: Forty-two healthy adults of both sexes, divided into male (MG) and female groups (FG), were provided respiratory questionnaires. Subsequently, the pulmonary function test was conducted to evaluate the ventilatory parameters of forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), and FEV1/FVC ratio in the sitting (S), dorsal decubitus (DD), right lateral decubitus (RLD), and left lateral decubitus (LLD) positions. A comparison of the parametric data was performed via one-way analysis of variance followed by Tukey post-hoc tests. Correlations between the S position variables along with the other positions were evaluated using the Pearson test. Results: The mean and standard error for the FVC values of the MG at positions DD (4.3 ± 0.7/L), RLD (4.1 ± 0.6/L) and LLD (4.1 ± 0.6/L) were lower when compared to S (5.05 ± 0.6 L). There was a strong positive correlation between the values of FVC, FEV1, and FEV1/FVC in the S position compared to other positions analyzed in both groups. Conclusion: Body positioning altered the parameters of the pulmonary function test in healthy adults. Resumo Introdução: A prova de função pulmonar, ou espirometria, é um teste validado e reconhecido mundialmente que contribui para o diagnóstico, estadiamento e acompanhamento longitudinal das doenças pulmonares. O exame é mais frequentemente realizado na posição sentada na prática clínica; portanto, não há valores previstos para seu desempenho em outras posições, como em decúbitos diferentes. Objetivo: O presente estudo teve como objetivo avaliar os efeitos da posição nos resultados dos testes de função pulmonar em adultos saudáveis. Métodos: Quarenta e dois adultos saudáveis de ambos os sexos, divididos nos grupos masculino (GM) e feminino (GF), receberam questionários respiratórios. Posteriormente, realizou-se o teste de função pulmonar para avaliar os parâmetros ventilatórios de capacidade vital forçada (CVF), volume expiratório forçado no primeiro segundo (VEF1) e relação VEF1/CVF nas posições sentada (S), decúbito dorsal (DD), decúbito lateral direito (DLD) e decúbito lateral esquerdo (DLE). A comparação dos dados paramétricos foi realizada por meio de análise de variância unidirecional seguida do pós-teste de Tukey. As correlações entre as variáveis da posição S com as demais posições foram avaliadas por meio do teste de Pearson. Resultados: A média e o erro padrão dos valores de CVF do MG nas posições DD (4,3 ± 0,7/L), DLD (4,1 ± 0,6/L) e DLE (4,1 ± 0,6/L) foram menores quando comparados com S (5,05 ± 0,6 L). Houve forte correlação positiva entre os valores de CVF, VEF1 e VEF1/CVF na posição S em relação às demais posições analisadas em ambos os grupos. Conclusão: O posicionamento corporal alterou os parâmetros do teste de função pulmonar em adultos saudáveis.

Chronic obstructive pulmonary disease (COPD) is the major cause of morbidity and mortality worldwide, and cigarette smoke is a key factor in the development of COPD. Thus, the development of effective therapies to prevent the advancement of COPD has become increasingly essential. We hypothesized that quercetin protects lungs in mice exposed to long-term cigarette smoke. Thirty-five C57BL/6 mice were exposed to cigarette smoke (12 cigarettes per day) for 60 days and pretreated with 10 mg/kg/day of quercetin via orogastric gavage. After the experimental protocol, the animals were euthanized and samples were collected for histopathological, antioxidant defense, oxidative stress and inflammatory analysis. The animals exposed to cigarette smoke showed an increase in respiratory rate and hematological parameters, cell influx into the airways, oxidative damage and inflammatory mediators, besides presenting with alterations in the pulmonary histoarchitecture. The animals receiving 10 mg/kg/day of quercetin that were exposed to cigarette smoke presented a reduction in cellular influx, less oxidative damage, reduction in cytokine levels, improvement in the histological pattern and improvement in pulmonary emphysema compared to the group that was only exposed to cigarette smoke. These results suggest that quercetin may be an agent in preventing pulmonary emphysema induced by cigarette smoke.

The present study aimed to evaluate the effects of maternal protein restriction during pregnancy on the lungs of 1-d and 31-d old offspring of C57BL/6 mice.The C57BL/6 mice (8-10 wk) were used for breeding. After pregnancy confirmation, female mice were randomly divided into a control group (CG) receiving a standard diet (22% protein) and a protein-restriction group (PRG) receiving a low-protein diet (6% protein). In the low-protein diet, protein was replaced by carbohydrate. After parturition, female mice that received the low-protein diet were fed the standard diet. Male offspring were euthanized 1 d and 31 d after birth for subsequent analysis. We evaluated the effects of a protein-restricted diet during gestation in pulmonary organogenesis, lung oxidative stress, and pulmonary inflammatory response of the offspring.PRG mice 1 d after birth showed lower body and lung mass, length, relative mass, lung density, and erythrocyte count compared with CG mice. There was an increase in alveolar airspace density and a higher mean linear intercept (Lm), greater oxidative damage, and inflammation in PRG mice compared with CG mice. At 31 d after birth, PRG mice had lower body mass, length, and lung mass values compared with CG mice. PRG mice showed greater recruitment of inflammatory cells to the airways. In addition, there was increased collagen deposition in the lungs, altered inflammatory mediators, and greater oxidative damage compared with CG mice.Protein restriction during pregnancy reduces the body weight of offspring and promotes inflammation and oxidative stress, resulting in a simplification of the lung structure.

Bronchial obstruction, caused by retained secretions, is often treated by the administration of mucoactive agents including distilled water, saline, hypertonic saline, and sodium bicarbonate. However, the inflammatory effect of these solutions on the lungs remains unclear. This study evaluated the instillation effects of different solutions on oxidative stress and lung inflammatory response in C57BL/6 mice. Fifty C57BL/6 mice were divided into 5 groups: control (CG); distilled water (DWG), hypertonic saline (HSG), saline (SG) and sodium bicarbonate (SBG). CG was exposed to ambient air while DWG, HSG, SG and SBG had 50 μl of respective solutions administered intranasally for 5 consecutive days. Twenty-four hours after the last intranasal instillation, all animals were euthanized for subsequent analysis. All solutions promoted increased recruitment of inflammatory cells to the lung compared to controls. Superoxide dismutase activity was lower in HSG compared to all other groups; catalase activity was reduced in SG, while it increased in SBG and DWG compared to CG. Finally, there was an increase in the inflammatory markers TNF-α, CCL2 and IFN-γ in DWG compared to CG, SG and HSG. In conclusions, the intranasal instillation of different solutions promotes redox imbalance and inflammation on lungs of adult mice.

Mechanical ventilation is an essential supportive therapy in the treatment of critical patients, and it aims to maintain adequate gas exchange; however, it can also contribute to inflammation and oxidative stress, thus leading to lung injury. We tested the hypothesis that exogenous surfactant administration will be protective against ventilator-induced lung injury in adult healthy Wistar rats both because of its anti-inflammatory properties as well as its role in preventing alveolar collapse at end-expiration. Thus, the effect of intranasal instillation of a bovine exogenous surfactant was tested in Wistar rats submitted to mechanical ventilation. The animals were divided into four groups: (1) CONTROL; (2) SURFACTANT; (3) Mechanical ventilation (MV); (4) MV with pre-treatment with surfactant (MVSURFACTANT). The MV and MVSURFACTANT were submitted to MV with high tidal volume (12 mL/kg) for 1 h. After the experimental protocol, all animals were euthanized and the arterial blood, bronchoalveolar lavage fluid and lungs were collected for biochemical, immunoenzymatic assay, arterial blood gases, and morphometric analyzes. The Wistar rats that received exogenous surfactant (Survanta®) by intranasal instillation before MV demonstrated reduced levels of leukocytes, inflammatory biomarkers such as CCL2, IL-1, IL-6 and TNF-α. Furthermore, it prevented oxidative damage by reducing lipid peroxidation and protein carbonylation as well as histological pattern changes of pulmonary parenchyma. Our data indicate that exogenous surfactant attenuated lung inflammation and redox imbalance induced by mechanical ventilation in healthy adult rats suggesting a preventive effect on ventilator-induced lung injury.

This study aimed to analyze the effects of volume-controlled ventilation (VCV) and pressure-controlled ventilation (PCV) modes in female Wistar rats. 18 Wistar female adult rats were divided into three groups: control (CG), pressure-controlled ventilation (PCVG), and volume-controlled ventilation (VCVG). PCVG and VCVG were submitted to MV for one hour with a tidal volume (TV) of 8 mL/Kg, respiratory rate of 80 breaths/min, and positive end-expiratory pressure of 0 cmH2O. At the end of the experiment, all animals were euthanized. The neutrophils and lymphocytes influx to lung were higher in VCVG and PCVG compared to CG. The activities of superoxide dismutase, catalase and myeloperoxidase were higher in PCVG compared to CG. There was an increase in lipid peroxidation and protein oxidation in PCVG compared to CG. The levels of CCL3 and CCL5 were higher in PCVG compared to CG. In conclusions, the PCV mode promoted structural changes in the lung parenchyma, redox imbalance and inflammation in healthy adult female rats submitted to MV.

Purpose: Aluminum is the third most abundant metal in the earth's crust and is widely used in industry. Chronic contact with aluminum results in a reduction in the activity of electron transport chain complexes, leading to excessive production of reactive oxygen species (ROS) and oxidative stress. This study aimed to evaluate the effects of short-term exposure of aluminum hydroxide on oxidative stress and pulmonary inflammatory response.Materials and methods: Male BALB/c mice were divided into three groups: control group (CG); phosphate buffered saline group (PBSG) and aluminum hydroxide group (AHG). CG was exposed to ambient air, while PBSG and AHG were exposed to PBS or aluminum hydroxide solutions via nebulization, three times per day for five consecutive days. Twenty-four hours after the last exposure, all animals were euthanized for subsequent analysis.Results: Exposure to aluminum hydroxide in the blood resulted in lower platelet levels, higher neutrophils, and lower monocytes compared to CG and PBSG. Aluminum hydroxide promoted the recruitment of inflammatory cells to the lung. Macrophage, neutrophil and lymphocyte counts were higher in AHG compared to CG and PBSG. Protein oxidation and superoxide dismutase activity were higher, while catalase activity and reduced and oxidizes glutathione ratio in AHG were lower compared to CG and PBSG. Furthermore, there was an increase in the inflammatory markers CCL2 and IFN-γ in AHG compared to CG and PBSG.Conclusion: In conclusion, short-term nebulization with aluminum hydroxide induces the influx of inflammatory cells and oxidative stress in adult BALB/c mice.

Cigarette smoke is highly toxic, and it can promote increased production of reactive species and inflammatory response and leads to liver diseases. Quercetin is a flavonoid that displays antioxidant and anti-inflammatory activities in liver diseases. This study aimed at evaluating the protective effects of quercetin on livers from mice exposed to long-term cigarette smoke exposure. Male C57BL/6 mice were divided into five groups: control (CG), vehicle (VG), quercetin (QG), cigarette smoke (CSG), quercetin, and cigarette smoke (QCSG). CSG and QCSG were exposed to cigarette smoke for sixty consecutive days; at the end of the exposures, all animals were euthanized. Mice that received quercetin daily and were exposed to cigarette smoke showed a reduced influx of inflammatory cells, oxidative stress, inflammatory reaction, and histopathological changes in the liver, compared to CSG. These results suggest that quercetin may be an effective adjuvant for treating damage to the liver due to cigarette smoke exposure.

Angiotensin-(1-7) [Ang-(1-7)], a peptide of the renin-angiotensin system, has anti-inflammatory, anti-fibrotic and antiproliferative effects in acute or chronic inflammatory disease of respiratory system. In this study, we evaluated the effect of treatment with Ang-(1-7) on pulmonary tissue damage and behavior of mice submitted to experimental model of elastase-induced pulmonary emphysema (PE). Initially, male C57BL/6 mice were randomly assigned into two main groups: control (CTRL) and PE. In the PE group, the animals received three intratracheal instillations of pancreatic porcine elastase (PPE) at 1-week intervals (0.2 IU in 50 μL of saline). The CTRL group received the same volume of saline solution (50 μL). Twenty-four hours after the last instillation, animals of the PE group were randomly divided into two groups: PE and PE + Ang-(1-7). The PE + Ang-(1-7) group was treated with 60 μg/kg of Ang-(1-7) and 92 μg kg of HPβCD in gavage distilled water, 100 μl. The CTRL and PE groups were treated with vehicle (HPβCD- 92 μg/kg in distilled water per gavage, 100 μl), orally daily for 3 weeks. On the 19th day of treatment, all groups were tested in relation to locomotor activity and exploratory behavior. After 48 h, the animals were euthanized and lungs were collected. The animals of PE group presented rupture of alveolar walls and consequently reduction of alveolar tissue area. Treatment with Ang-(1-7) partially restored the alveolar tissue area. The PE reduced the locomotor activity and the exploratory behavior of the mice in relation to the control group. Treatment with Ang-(1-7) attenuated this change. In addition, it was observed that Ang-(1-7) reduced lung levels of IL-1β and increased levels of IL-10. These results show an anti-inflammatory effect of Ang-(1-7), inducing the return of pulmonary homeostasis and attenuation of the behavioral changes in experimental model of PE by elastase.

An ethnopharmacological survey indicates that the genus Byrsonima has some medicinal species that are commonly found in the Brazilian Cerrado and has been used as an anti-inflammatory and for gastroduodenal disorders. The aim of this study was to evaluate the anti-inflammatory and antioxidant activity along with qualitative chemical characterization of the methanolic extract of the leaves of Byrsonima verbascifolia (BvME) obtained by exhaustive percolation. The data from the chemical analyses by liquid chromatography-mass spectrometry led to tentative identification of 42 compounds belonging to proanthocyanidins, galloyl quinic acid derivatives, flavonoids, and triterpene glycoside derivatives. BvME contain flavonoids and show an antioxidative activity. The methanolic extract administered intraperitoneally at doses of 50, 100, or 300 mg/kg showed a significant reduction in paw edema and modulated the neutrophil influx in a mouse model. Furthermore, the anti-edematogenic activity of the extract provided in smaller doses (12.5 and 25 mg/kg) was also demonstrated in a mouse paw edema model. The extract inhibited NO production by macrophages induced by lipopolysaccharide. We presume that the anti-inflammatory effects of BvME are due to a combination of compounds present in B. verbascifolia, including catechins (procyanidins), flavonoids, and triterpene glycosides and that these anti-inflammatory actions should be mediated, at least partly, through the inhibition of NO production. This study supports and validates the ethnopharmacological uses of B. verbascifolia as an anti-inflammatory.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Os receptores ativados por proteases (do inglês, proteinase-activated receptor, PAR) fazem parte de uma família de receptores acoplados a proteína G, que são ativados principalmente pela clivagem proteolítica de uma sequência específica de aminoácidos localizados em seu domínio N-terminal. Sua ativação vem sendo associada à regulação de diferentes fenômenos inflamatórios. PAR-2 é um receptor com potencial para regular a inflamação alérgica uma vez que está presente em uma grande variedade das células das vias aéreas como, por exemplo, células epiteliais e macrófagos alveolares. O objetivo do presente estudo foi avaliar a importância da ativação do PAR-2 na produção de mediadores inflamatórios e no recrutamento de leucócitos em modelos experimentais de asma induzidos por ovalbumina (OVA). Os protocolos experimentais foram aprovados pela comissão de ética no uso de animais (CEUA/UFMG, nº348/2014). A inflamação pulmonar alérgica foi induzida por OVA (10μg, via intranasal) em camundongos Balb/c fêmeas (8-10 semanas) previamente iminizados para o antígeno, em dois modelos experimentais diferindo entre si o número de desafios intranasais, três ou seis. Dentre o período de 30min à 72h após último desafio com o alérgeno, os camundongos foram sacrificados, sendo em seguida coletados os lavados broncoalveolares (LBA) e feita a remoção dos pulmões. Para avaliar a participação do PAR-2 na asma experimental, os camundongos imunizados foram pré-tratados com o antagonista seletivo de PAR-2, ENMD1068 (0,1–1,0 mg/ kg, via intraperitoneal) 1 hora antes da administração intranasal de OVA. O bloqueio de PAR-2 promoveu efeito considerável no controle da inflamação pulmonar alérgica em ambos os modelos de asma, reduzindo a presença de neutrófilos, eosinófilos e células mononucleares no LBA dos camundongos quando comparado aqueles desafiados com OVA e pré-tratados apenas com PBS. Especificamente no modelo de três exposições à OVA, o antagonismo do receptor reduziu a expressão protéica de PAR-2 em leucócitos alveolares, a produção de CXCL1, IL-6 e CCL5 no LBA e de IL-1β no pulmão, a permeabilidade vascular no tecido pulmonar e a população de macrófagos pulmonares com fenótipo M2, além de aumentar os níveis de IL-10 no LBA quando comparado ao grupo OVA. No modelo de seis exposições à OVA, o bloqueio de PAR- 2 conduziu à redução nas atividades de peroxidase eosinofílica (EPO) e mieloperoxidase (MPO) no parênquima pulmonar, no extravasamento de proteínas para o LBA, evitou a perda da complacência pulmonar induzida pela OVA, diminuiu a resistência ao fluxo de ar basal e em reposta à provocação com metacolina, além de diminuir a produção de muco e deposição de colágeno em tecido pulmonar, quando comparados aos animais desafiados com OVA. Nossos resultados demonstram que há modulação da resposta imunológica via ativação de PAR-2 na asma experimental em camundongos BALB/c nos diferentes modelos avaliados. Além disso, o bloqueio da ação de proteases endógenas ativando PAR-2 em camundongos tratados com o antagonista seletivo desempenhou um papel importante no controle da inflamação pulmonar alérgica, sugerindo que o bloqueio de PAR-2 pode ser útil como uma nova abordagem farmacológica em relação ao tratamento de doenças respiratórias das vias aéreas. Proteinase-activated receptors (PAR) are part of a family of G-protein-coupled receptors that are activated via proteolytic cleavage of a specific sequence of amino acids in N-terminal portion, and their activation has been implicated in the regulation of inflammation. PAR-2 has been implicated in mediating allergic airway inflammation once is expressed by many cells in the airways as well as in epithelial and alveolar macrophages. The aim of this study was to study the role of PAR-2 activation in the production of inflammatory mediators and on the recruitment of leukocytes in experimental ovalbumin (OVA)-induced asthma models. All experimental protocols were approved by local UFMG Ethics’s Committee for Animal Use - certificate number 348/2014. Allergic lung inflammation was induced in sensitized BALB/c mice (female, 8-10 weeks) through the intranasal instillation of ovalbumin (OVA, 10μg) in two distinct experimental models differing in relation to the number of intranasal challenges (3 or 6 challenges). Bronchoalveolar lavage (BAL) was performed and lungs were obtained in different intervals of time, from 30 min to 72 h after challenging. To evaluate the participation of PAR-2 in experimental asthma, sensitized mice were pretreated with selective PAR-2 antagonist (ENMD1068, 0.1-1.0 mg / kg, intraperitoneal) 1 hour before intranasal instillation of OVA. The blockade of the action of PAR-2 activating endogenous proteases in PAR-2 antagonist-treated mice, played an important role on the control of allergic respiratory inflammation in both asthma’s models. Overall, pretreatment with PAR-2 antagonist ENMD1068 inhibited the recruitment of neutrophils, eosinophils, mononuclear cells to LBA when compared to OVA-treated mice. After OVA instillation (3 intranasal challenge model), PAR-2 blockade reduced PAR-2 protein expression in leukocytes, CXCL1, IL-6 and CCL5 production in BAL and IL-1β releasing into the lung, vascular permeability in lung and M2 lung macrophages population. In addition, PAR-2 blockade increased IL-10 levels in BAL when compared to OVA group. Six intranasal challenge model also promotes leukocyte infiltrating into BAL in a PAR2-dependent manner. PAR-2 blockade impairs eosinophil peroxidase (EPO) and myeloperoxidase (MPO) activity at the parenchyma lung, proteins extravasation into BAL, reduces the loss of dynamic pulmonary compliance and the lung resistance in response to methacholine, as well as mucus production and collagen deposition when compared to OVA-challenged. In the present study we demonstrated a role for PAR-2 activation in the modulation of essential phenomena leading to allergic asthma in BALB/c mice both models. In addition, the blockade of the action of PAR-2 activating endogenous proteases in PAR-2 antagonist-treated mice played an important role on the control of allergic respiratory inflammation, suggesting that PAR-2 blockade may be useful as a new pharmacological approach regarding the treatment of airways allergic diseases.

Annona crassiflora Mart., popularly known as “Araticum”, is a native tree of the Brazilian Cerrado used in folk medicine for treatment of pain and inflammatory diseases. We proposed to analyze analgesic and anti-inflammatory properties of the filtrate (F1) and the precipitate (F2) of the hydroalcoholic fraction from the leaves of Annona crassiflora Mart. in mice. Swiss mice were submitted to formalin-induced nociception test and tail-flick reflex test, to assess antinociceptive properties, and to the rota-rod test, for motor performance analyses. To evaluate anti-inflammatory properties, F1 and F2 were orally administered 1 h prior to the intrathoracic injection of carrageenan, zymosan, LPS, CXCL8, or vehicle in Balb/c mice and neutrophil infiltration was evaluated 4 h after injection. F1 and F2 reduced the licking time in the second phase of formalin-induced nociception test, but only F2 showed a dose-dependent response. Neither F1 nor F2 reduced the latency time in the tail-flick reflex test. In addition, motor performance alteration was not observed in F1- or F2-treated mice. F2 treatment significantly inhibited the neutrophilia induced by carrageenan, LPS, or CXCL8, but not zymosan. The experimental data demonstrated that hydroalcoholic fractions of Annona crassiflora Mart. leaves have remarkable anti-inflammatory and antinociceptive activities.

Although proteinase-activated receptor (PAR)-2 has been implicated in inflammatory diseases, its role in regulating eosinophil recruitment in response to chemoattractants remains unclear. Here, we investigated the role of PAR-2 and PAR-2-activating Mast Cell (MC) tryptase on chemokine C–C motif ligand (CCL)11- and antigen-induced eosinophil recruitment to the pleural cavity of BALB/c mice. The PAR-2-activating peptide H–Ser–Leu–Ile–Gly–Arg–Leu–NH2 (SLIGRL–NH2) induced eosinophil recruitment whereas PAR-2 blockade inhibited ovalbumin (OVA)- or CCL11-induced eosinophil recruitment. Moreover, OVA and CCL11 induced PAR-2 expression in pleural leukocytes, and the MC tryptase inhibitor APC 366 ([N-(1-hydroxy-2-napthoyl)- l -arginyl- l -prolinamide hydrochloride]) abolished CCL11-induced eosinophil recruitment. These results suggest a pro inflammatory effect of PAR-2 and support a role for MC tryptase mediating eosinophil migration via PAR-2 signaling. Taken together, our results suggest that PAR-2 activation through endogenous MC tryptase activity could be required, at least partially, to mediate CCL11-induced eosinophil migration.

Byrsonima verbascifolia (Malpighiaceae), commonly known as 'murici', is used in folk medicine, for example, in the treatment of inflammation. The anti-inflammatory activity of the butanolic fraction of B. verbascifolia leaves (BvBF) was previously reported by our group, and the present study was designed to evaluate their antinociceptive effects. BvBF (25, 50, and 100 mg/kg) administered intraperitoneally (i.p.) inhibited acetic acid induced abdominal writhing. In the formalin test, BvBF (10, 30 and 100 mg/kg, i.p.) caused a reduction in licking time in both the neurogenic and inflammatory phases. Moreover, we demonstrated that BvBF (30 and 100 mg/kg, i.p.) caused an increase in the latency to response in the hot-plate test. These results demonstrate that BvBF possesses marked peripheral and central antinociceptive activities. Pre-treatment with the non-selective receptor antagonist naloxone (5 mg/kg, i.p.) abolished the antinociceptive effects of BvBF (100 mg/kg, i.p.) in the neurogenic phase of the formalin and hot-plate tests. The anti-inflammatory activity of BvBF (previously reported) as well as the participation of the opioidergic system seems to be responsible, at least in part, for these antinociceptive effects. Finally, BvBF at the doses investigated (25, 50 and 100 mg/Kg) did not cause any toxicity signals, showing that the antinociceptive activity is devoid of sedative and hypomotility effects.

The activation of proteinase-activated receptors (PARs) has been implicated in the development of important hallmarks of inflammation, including in vivo leukocyte recruitment; however, its role in the regulation of leukocyte migration in response to inflammatory stimuli has not been elucidated until now. Here, we examined the effects of the PAR4 antagonist YPGKF-NH 2 (tcY-NH2) on neutrophil recruitment in experimentally induced inflammation. BALB/c mice were intrapleurally injected with tcY-NH2 (40 ng/kg) prior to intrapleural injection of carrageenan (Cg) or neutrophil chemoattractant CXCL8; the number of infiltrating neutrophils was evaluated after 4 h, and KC production was assessed at different times after Cg injection. Neutrophil adhesion and rolling cells were studied using a brain circulation preparation 4 h after the Cg or CXCL8 challenge in tcY-NH2-treated mice. PAR4 blockade inhibited CXCL8- and Cg-induced neutrophil migration into the pleural cavity of BALB/c mice and reduced neutrophil rolling and adherence. Surprisingly, PAR4 blockade increased the level of KC in response to carrageenan. These results demonstrated that PAR4 blockade impairs neutrophil migration in vivo, suggesting that PAR4 plays an important role in the regulation of inflammation, at least in part because of its ability to inhibit the actions of the neutrophil chemoattractant CXCL8.

Proteinase-activated receptor (PAR) 2 has been implicated in eosinophil migration. Mast cell (MC) tryptase has been similarly implicated in allergic diseases through the activation of PAR-2, but the role of this receptor in MC tryptase-induced inflammation is not well elucidated. This study aims to investigate the ability of MC tryptase or PAR-2 activating peptide (SLIGRL-NH2) to induce eosinophil recruitment to the pleural cavity of mice. Mast cell tryptase-injected mice were pretreated with PAR-2 antagonist ENMD-1068. Mice injected with SLIGRL-NH2 were pretreated with mast cell tryptase inhibitor APC 366, and eosinophil migration into the pleural cavity and PAR-2 expression was analyzed after 24 or 48 h. SLIGRL-NH2-induced eosinophil recruitment was inhibited by APC 366, and MC tryptase-induced eosinophil recruitment was abolished by ENMD-1068. MC tryptase induced PAR-2 expression on pleural eosinophils. Our results demonstrate a key role for PAR-2 in mediating eosinophil recruitment in MC tryptase-induced pleurisy in mice. The ability of MC tryptase to inducing PAR-2 expression on eosinophils corroborates the relevance of MC tryptase and PAR-2 on modulating eosinophil migration.

As plantas medicinais se destacam como grandes fontes para o desenvolvimento de medicamentos e a selecao de especies vegetais para a realizacao de estudos farmacologicos e fitoquimicos pode ser baseada na abordagem etnofarmacologica. Realizamos a analise fitoquimica preliminar da fracao butanolica obtida das folhas de Byrsonima verbascifolia e avaliamos sua atividade antiinflamatoria atraves dos modelos edema de pata e pleurisia induzidos pela carragenina. A analise fitoquimica caracterizou a presenca de compostos fenolicos como os derivados flavonicos. A atividade antiinflamatoria da fracao foi evidenciada pela reducao do edema de pata e diminuicao da migracao dos neutrofilos para a cavidade pleural. A fracao butanolica na dose de 12,50 mg/Kg reduziu 84,21% (P