Severe Asthma: Updated Therapy Approach Based on Phenotype and Biomarker

Asthma is responsible for considerable global morbidity and health-care costs affecting over 300 million people worldwide. This illness is a heterogeneous condition characterized by chronic airway inflammation and pulmonary tissue remodeling resulting in a variety of clinical manifestations and treatment responses. Recent studies have shown an increasing appreciation of heterogeneity in asthma based on molecular phenotyping, biomarkers, and differential responses to therapies. In terms of asthma classification, perhaps the most important distinction to make is whether the patient has evidence of an eosinophilic inflammatory process characterized by type 2 immune response (Th2) or not. Therefore, personalized therapies to asthmatic patients just will be a reality by identifying and characterizing biomarkers. This review approaches the advances in diagnoses and management of asthma and severe asthma and highlights those with difficult-to-treat asthma based on each phenotype and biomarkers, to assist in the optimization of conventional therapy and to guide the use of targeted therapies

Estudo da atividade ansiolítica e imunomoduladora do Gama-terpineno

Inflammation is generated in response to noxious stimuli, such as pathogens, and may be associated with an acute pathogenesis or progress to a chronic process. Gamma-terpinene (GT) is a monoterpene that, in previous studies, presented antiinflammatory action in acute inflammatory processes. The present work aimed to elucidate the anti-inflammatory and anxiolytic mechanisms of GT in murine models of acute (Acute Pulmonary Injury - ALI) and chronic inflammation (rhinitis and asthma). For this, BALB/c mice were challenged with lipopolysaccharide (LPS) to induce ALI and were treated one hour post challenge with GT (12.5, 25 and 50 mg/kg) for three days. After 72 h the animals were euthanized for collection of bronchoalveolar lavage fluid (BALF). For murine models of rhinitis and asthma, BALB/c mice were sensitized and challenged with ovalbumin (OVA) and were treated with GT (12.5; 25 and 50 mg/kg) or standard drug (dexamethasone 2 mg/kg) 1 h before each challenge with OVA. After the last challenge, animals were submitted to clinical symptoms analysis of rhinitis and anxiety and immunological tests in the asthma model. In the ALI protocol, animals GT-treated (50 mg/kg) presented, in nasal fluid lavage (NALF), a decrease in the migration of total and differential cells (neutrophils and mononuclear cells), total protein concentration, as well as lung weight, when compared with OVA group. Histopathological studies showed that treatment with GT50 decreased the cellular infiltrate in the alveolar and vascular regions, edema and hemorrhage. In immunological parameters, GT50 treatment presented a decrease in inflammatory cytokines, such as IL-1β, IL-6 and TNF-α, as well as decreases in the expression of the TLR4 receptor, p38 MAPKinase and p65NFkB. In rhinitis protocol, treatment with GT50 showed a decrease in clinical symptoms, such as nasal friction and sneezing, in addition to a decrease in BALF of total and differential cells, mainly eosinophils. Histopathological analyses of the nasal cavity, the treatment with GT50 promoted a decrease in cell infiltrate, mucus secretion and mast cell number. The treatment with GT50 in asthma protocol showed a significantly decrease in anxiety rates compared to OVA animals (sick), effect similar to diazepam (1 mg/kg), presenting an anxiolytic effect. Treatment with GT50 also decreased c-fos expression in brain areas such as the hypothalamic paraventricular nucleus (PVN) and central nucleus of the amygdala (CeA) when compared to the OVA group. In the histological analyzes there was a decrease in cell infiltration and mucus secretion in the lung tissue. In addition, treatment with GT50 decreased immunological parameters such as: IgE-OVAspecific levels; migration of total and differential cells mainly eosinophil in BALF and modulation of the Th2 response by inducing both Th1 profile by secretion of IFN cytokine and regulatory profile, by secretion of cytokine IL-10. Treatment with GT50 was also able to decrease Th2 profile cytokines, such as IL-4 and IL-13, in addition to decreasing the activation of p38 MAPquinase and p65NFkB receptor expression. These data indicate that GT has both anxiolytic and anti-inflammatory properties in acute and chronic processes by inhibition of MAP kinasep38 and the transcription factor NFκB.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqA inflamação é gerada em resposta a estímulos nocivos, tais como agentes patogênicos, podendo estar associada a uma patogênese aguda ou evoluir para um processo crônico. O gama-terpineno (GT) é um monoterpeno que, em estudos prévios, apresentou ação anti-inflamatória em processos inflamatórios agudos. O presente trabalho teve como objetivo elucidar os mecanismos anti-inflamatórios e ansiolíticos do GT em modelos murinos de inflamação aguda - Lesão Pulmonar Aguda (LPA) e inflamação crônica - rinite e asma. Para tal, camundongos BALB/c foram desafiados com lipopolissacarídeo (LPS) para induzir a LPA e foram tratados, uma hora após o desafio, com GT (12,5; 25 e 50 mg/kg) por três dias. Após 72 h os animais foram eutanasiados para coleta do fluido do lavado broncoalveolar (BALF). Para os modelos murinos de rinite e asma, camundongos BALB/c foram sensibilizados e desafiados com ovalbumina (OVA) e foram tratados com GT (12,5; 25 e 50 mg/kg) ou droga padrão (dexametasona 2 mg/kg) 1 h antes de cada desafio com OVA. Após o último desafio, os animais foram submetidos à análise de sintomas clínicos da rinite e testes de ansiedade e imunológicos no modelo de asma. Na LPA, os animais tratados com GT (50 mg/kg) apresentaram diminuição, no lavado do fluido nasal (NALF), na migração das células totais, diferenciais (neutrófilos e células mononucleares), na concentração de proteínas totais, bem como no peso pulmonar quando comparados ao do grupo OVA (doente). Estudos histopatológicos mostraram que o tratamento com GT50 diminuiu o infiltrado celular nas regiões alveolares e vasculares, edema, hemorragia. Na análise de parâmetros imunológicos, houve diminuição das citocinas inflamatórias, IL-1β, IL-6 e TNF-α, bem como diminuição da expressão do receptor TLR4, da MAPquinase p38 e do p65NFkB. No protocolo de rinite, o tratamento com GT50 apresentou diminuição dos sintomas clínicos, como fricção nasal e espirros, além de diminuição no BALF de células totais e diferenciais, principalmente eosinófilos. Na histopatologia da cavidade nasal, o tratamento com GT50 promoveu diminuição nesta área: do infiltrado celular, da secreção de muco e do número de mastócitos. Na asma, o tratamento com GT50 diminuiu significativamente os índices de ansiedade em comparação com animais do grupo OVA (doente) de forma semelhantes ao diazepam (1 mg/kg), apresentando efeito ansiolítico. O tratamento com o GT50 também diminuiu a expressão de c-fos em áreas cerebrais como o núcleo paraventricular hipotalâmico (PVN) e núcleo central da amigdala (CeA) quando comparado com o grupo OVA. Nas análises histológicas houve diminuição do infiltrado e da secreção de muco no tecido pulmonar. Em adição, o tratamento com GT50 diminuiu parâmetros imunológicos como: níveis de IgE-OVA-específica; migração de células totais e diferenciais principalmente eosinófilo no BALF e modulação da resposta Th2 por induzir tanto a citocinas do perfil Th1 como IFN como do perfil regulatório IL-10. O tratamento com GT50 também foi capaz de diminuir as citocinas Th2, como a IL-4 e IL-13 além de diminuir a ativação da MAPquinase p38 e a expressão do p65NFkB. Esses dados indicam que o GT possui ambas as propriedades: ansiolítica e anti-inflamatória em processos agudos e crônicos pela inibição da MAP quinasep38 e do fator de transcrição NFkB

Anti-inflammatory activity and acute toxicity studies of hydroalcoholic extract of Herissantia tiubae

ABSTRACT Hydroalcoholic extract of aerial parts of Herissantia tiubae (K. Schum.) Brizicky, Malvaceae, was evaluated in experimental models of inflammation and toxicity. For toxicity assays, male and female Swiss mice were orally treated with hydroalcoholic extract of H. tiubae (2000 mg/kg) and analyzed by consumption of water and food, body weight, mortality and rates of major organ weights, as well as biochemical and hematological indexes. For anti-inflammatory effect, phlogistic agents such as carrageenan or acetic acid were used to evaluate paw edema, cell migration and cytokine production. It was also investigated the hydroalcoholic extract of H. tiubae in RAW 264.7 macrophage lineage by nitric oxide and cytokine productions. Swiss mice treated with hydroalcoholic extract of H. tiubae showed low toxicity and (50 or 100 mg/kg) was able to reduce significantly (p < 0.01, p < 0.001) polymorphonuclear cell migration, TNF-&#945; and IL-1&#946; production in the carrageenan-induced peritonitis. However the hydroalcoholic extract of H. tiubae (50, 100 or 200 mg/kg) did not reduce carrageenan-induced paw edema. Additionally, hydroalcoholic extract of H. tiubae did not present cytotoxicity at concentrations of 6.25, 12.5, 25 or 50 µg/ml but induced significantly decrease of NO, TNF-&#945; and IL-6 production in macrophage lineage. This study suggests that hydroalcoholic extract of H. tiubae has anti-inflammatory activity by inhibiting cell migration mainly by decreasing the inflammatory cytokine levels at the inflamed site independently of the anti-edematogenic effect

Flavonoides isolados de Sida santaremnensis H. Monteiro (“Guanxuma”) e avaliação das atividades biológicas

Introduction: Sida santaremnensis H. Monteiro (Malvaceae) is a plant popularly known as "vassourinha" or "guanxuma" that has been described as vasorelaxant, antiulcerogenic, antinociceptive and antiedematogenic. Objective: To contribute with the phytochemical and pharmacological profile of Sida santaremnensis through the isolation, purification and determination of chemical constituents of this plant, as well as through the evaluation of the anti-inflammatory and antitumor activity and leishmanicidal effect of the isolated constituent in a greater amount. Methods: The isolation of substances from the plant herein studied was performed with column chromatographic and analytical thin-layer methods and structural determination made by spectroscopic methods, such as Nuclear Magnetic Resonance, Hydrogen and Carbon 13, and comparisons with data from the literature. To assess pharmacological activities, cell viability tests, determination of nitric oxide levels, leishmanicidal activity, among others, were performed. Results: Two flavonoids from S. santaremnensis were obtained, kaempferol (S-1) and kaepferol 3-O-β-D-glycosyl-6’’-α-L-rhamnoside (S-2), the latter in a greater amount. The evaluation of the antitumor activity of the glycosylated flavonoid demonstrated that it does not present hemolytic activity against the human promyelocytic leukemia cell line (HL-60), and that it presented weak leishmanicidal activity. The immunopharmacological evaluation of kaempferol 3-O-β-D-glucosyl-6’’-α-L-rhamnoside revealed that it presents a possible anti-inflammatory action related to the inhibition of the production of nitrite by LPS-stimulated macrophages. Conclusion: These data demonstrate that kaempferol 3-O-β-D-glucosyl-6’’-α-L-rhamnoside has low toxicity, in addition to antileishmania, anti-inflammatory and immunomodulatory properties, which makes it a therapeutic potential for infectious and inflammatory diseases mediated by macrophages.Sida santaremnensis H. Monteiro (Malvaceae) é uma planta conhecida popularmente como "vassourinha" ou "guanxuma", que vem sendo descrita como vasorelaxante, antiulcerogênica, antinociceptiva e antiedematogênica. Objetivou-se contribuir com o estudo fitoquímico e farmacológico de Sida santaremnensis a partir do isolamento e determinação estrutural de constituintes químicos dessa planta, bem como pela avaliação da atividade anti-inflamatória e antitumoral do constituinte isolado em maior quantidade. O isolamento de substâncias da planta estudada foi realizado por métodos cromatográficos em coluna e em camada delgada analítica e a determinação estrutural feita por métodos espectroscópicos, como a Ressonância Magnética Nuclear de Hidrogênio e Carbono 13, e comparações com a literatura. Para avaliação das atividades farmacológicas foram realizados ensaio de viabilidade celular, determinação dos níveis de óxido nítrico, atividade leishmanicida, entre outros. Foram obtidos como resultados dois flavonoides de S. santaremnensis, o canferol (S-1) e canferol 3-O-β-D-glycosyl-6’’-α-L-ramnosídeo (S-2), este último em maior quantidade. A avaliação da atividade antitumoral do flavonoide glicosilado demonstrou que ele não apresenta atividade hemolítica nem citotóxica contra linhagem de células de leucemia promielocítica humana (HL-60), porém apresentou fraca atividade leishmanicida. A avaliação imunofarmacológica dessa mesma substância revelou uma possível ação anti-inflamatória relacionada à inibição da produção de nitrito por macrófagos estimulados com LPS. Conclui-se que estes dados demonstram que o canferol 3-O-β-D-glicosil-6’’-α-L-ramnosídeo apresenta baixa toxicidade, além de propriedades antileishmania, anti-inflamatória e imunomoduladora, o que o torna um potencial terapêutico para doenças infecciosas e inflamatórias mediadas por macrófagos