Bactericidal and anti-inflammatory effects of Moquilea tomentosa Benth. flavonoid-rich leaf extract.

BACKGROUND: Natural products are an important source of bioproducts with pharmacological properties. Here we investigate the components of leaves from M. tomentosa Benth. (Fritsch) (Chrysobalanaceae) and its effects on bacterial cell growth, biofilm production and macrophage activity. METHODS: The effect of the different leaf extracts against bacterial cell growth was performed using the microdilution method. The most active extract was analyzed by mass spectrometry, and its effect on bacterial biofilm production was evaluated on polystyrene plates. The extract effect on macrophage activity was tested in the RAW264.7 cell line, which was stimulated with different concentrations of the extract in the presence or absence of LPS. RESULTS: We show that the ethyl acetate (EtOAc) extract was the most effective against bacterial cell growth. EtOAc extract DI-ESI (-)MSn analysis showed the presence of a glycosylated flavonoid tentatively assigned as myricetin 3-O-xylosyl-rhamnoside (MW 596). Also, the EtOAc extract increased biofilm formation by S. aureus and inhibited cytokine and NO production induced by LPS in RAW macrophages. CONCLUSION: M. tomentosa flavonoid-enriched EtOAc extract presented a bactericidal and anti-inflammatory pharmacological potential.

Glutamine Therapy Reduces Inflammation and Extracellular Trap Release in Experimental Acute Respiratory Distress Syndrome of Pulmonary Origin.

The innate immune response plays an important role in the pathophysiology of acute respiratory distress syndrome (ARDS). Glutamine (Gln) decreases lung inflammation in experimental ARDS, but its impact on the formation of extracellular traps (ETs) in the lung is unknown. In a mouse model of endotoxin-induced pulmonary ARDS, the effects of Gln treatment on leukocyte counts and ET content in bronchoalveolar lavage fluid (BALF), inflammatory profile in lung tissue, and lung morphofunction were evaluated in vivo. Furthermore, ET formation, reactive oxygen species (ROS) production, glutathione peroxidase (GPx), and glutathione reductase (GR) activities were tested in vitro. Our in vivo results demonstrated that Gln treatment reduced ET release (as indicated by cell-free-DNA content and myeloperoxidase activity), decreased lung inflammation (reductions in interferon-γ and increases in interleukin-10 levels), and improved lung morpho-function (decreased static lung elastance and alveolar collapse) in comparison with ARDS animals treated with saline. Moreover, Gln reduced ET and ROS formation in BALF cells stimulated with lipopolysaccharide in vitro, but it did not alter GPx or GR activity. In this model of endotoxin-induced pulmonary ARDS, treatment with Gln reduced pulmonary functional and morphological impairment, inflammation, and ET release in the lung.

Eicosapentaenoic Acid Enhances the Effects of Mesenchymal Stromal Cell Therapy in Experimental Allergic Asthma.

Asthma is characterized by chronic lung inflammation and airway hyperresponsiveness. Despite recent advances in the understanding of its pathophysiology, asthma remains a major public health problem and, at present, there are no effective interventions capable of reversing airway remodeling. Mesenchymal stromal cell (MSC)-based therapy mitigates lung inflammation in experimental allergic asthma; however, its ability to reduce airway remodeling is limited. We aimed to investigate whether pre-treatment with eicosapentaenoic acid (EPA) potentiates the therapeutic properties of MSCs in experimental allergic asthma. Seventy-two C57BL/6 mice were used. House dust mite (HDM) extract was intranasally administered to induce severe allergic asthma in mice. Unstimulated or EPA-stimulated MSCs were administered intratracheally 24 h after final HDM challenge. Lung mechanics, histology, protein levels of biomarkers, and cellularity in bronchoalveolar lavage fluid (BALF), thymus, lymph nodes, and bone marrow were analyzed. Furthermore, the effects of EPA on lipid body formation and secretion of resolvin-D1 (RvD1), prostaglandin E2 (PGE2), interleukin (IL)-10, and transforming growth factor (TGF)-ß1 by MSCs were evaluated in vitro. EPA-stimulated MSCs, compared to unstimulated MSCs, yielded greater therapeutic effects by further reducing bronchoconstriction, alveolar collapse, total cell counts (in BALF, bone marrow, and lymph nodes), and collagen fiber content in airways, while increasing IL-10 levels in BALF and M2 macrophage counts in lungs. In conclusion, EPA potentiated MSC-based therapy in experimental allergic asthma, leading to increased secretion of pro-resolution and anti-inflammatory mediators (RvD1, PGE2, IL-10, and TGF-ß), modulation of macrophages toward an anti-inflammatory phenotype, and reduction in the remodeling process. Taken together, these modifications may explain the greater improvement in lung mechanics obtained. This may be a promising novel strategy to potentiate MSCs effects.

Human adipose tissue mesenchymal stromal cells and their extracellular vesicles act differentially on lung mechanics and inflammation in experimental allergic asthma.

BACKGROUND: Asthma is a chronic inflammatory disease that can be difficult to treat due to its complex pathophysiology. Most current drugs focus on controlling the inflammatory process, but are unable to revert the changes of tissue remodeling. Human mesenchymal stromal cells (MSCs) are effective at reducing inflammation and tissue remodeling; nevertheless, no study has evaluated the therapeutic effects of extracellular vesicles (EVs) obtained from human adipose tissue-derived MSCs (AD-MSC) on established airway remodeling in experimental allergic asthma. METHODS: C57BL/6 female mice were sensitized and challenged with ovalbumin (OVA). Control (CTRL) animals received saline solution using the same protocol. One day after the last challenge, each group received saline, 105 human AD-MSCs, or EVs (released by 105 AD-MSCs). Seven days after treatment, animals were anesthetized for lung function assessment and subsequently euthanized. Bronchoalveolar lavage fluid (BALF), lungs, thymus, and mediastinal lymph nodes were harvested for analysis of inflammation. Collagen fiber content of airways and lung parenchyma were also evaluated. RESULTS: In OVA animals, AD-MSCs and EVs acted differently on static lung elastance and on BALF regulatory T cells, CD3+CD4+ T cells, and pro-inflammatory mediators (interleukin [IL]-4, IL-5, IL-13, and eotaxin), but similarly reduced eosinophils in lung tissue, collagen fiber content in airways and lung parenchyma, levels of transforming growth factor-ß in lung tissue, and CD3+CD4+ T cell counts in the thymus. No significant changes were observed in total cell count or percentage of CD3+CD4+ T cells in the mediastinal lymph nodes. CONCLUSIONS: In this immunocompetent mouse model of allergic asthma, human AD-MSCs and EVs effectively reduced eosinophil counts in lung tissue and BALF and modulated airway remodeling, but their effects on T cells differed in lung and thymus. EVs may hold promise for asthma; however, further studies are required to elucidate the different mechanisms of action of AD-MSCs versus their EVs.

Medicação anti-inflamatória alternativa aos glicocorticóides para o tratamento da asma: papel dos modificadores de leucotrienos, estabilizadores de mastócitos e outras perspectivas / Anti-inflammatory medication beyond glucocorticoids for asthma therapy: role of leukotriene

A inalação de agentes anti-inflamatórios esteroidais, administrados isoladamente ou combinados a agonistas β2-adrenérgicos de longa duração, é a melhor opção disponível para o controle farmacológico da asma, embora efeitos adversose resistência aos glicocorticóides limitem seu benefício. Inibidores da síntese de leucotrienos, antagonistas do receptor CysLT1 e estabilizadores de mastócitos têm sido empregados, como monoterapias alternativas, no tratamento da asma branda e moderada, porém sem a mesma eficácia dos anti-inflamatórios esteroidais. Há evidências de que a combinação do antagonista CysLT1 e glicocorticóide inalado seja eficaz no tratamento de asmáticos graves, com diminuição da dosagem do agente esteroidal. Inibidores da enzima fosfodiesterase 4, bem como lidocaína e análogos não anestésicos da lidocaína, têm igualmente atraído atenção como opções terapêuticas no controle da asma. Esta revisão analisa avanços recentes no campo das alternativas ao uso dos glicocorticóides para regulação anti-inflamatória da asma

JMF2-1, a lidocaine derivative acting on airways spasm and lung allergic inflammation in rats.

BACKGROUND: Prior reports show that nebulized lidocaine might be an effective treatment for asthma. OBJECTIVE: We sought to determine the anti-inflammatory and spasmolytic effects of lidocaine and its analogue, JMF2-1, which we have synthesized for reduced local anesthetic activity. METHODS: Blockade of Na(+) currents was assayed in cultured GH(3) cells by using the patch-clamp technique, whereas anesthesia was assessed in a cutaneous pinching test in rats. Lidocaine and its analogue were nebulized into sensitized rats for evaluation of their effectiveness on airways spasm and inflammation induced by methacholine and allergen, respectively. Tissue histamine release and tracheal spasm triggered by allergen challenge in the absence and presence of these treatments were also examined in vitro. RESULTS: The 50% inhibitory concentration values for blockade of Na(+) currents after treatment with JMF2-1 (25.4 mM) was remarkably higher than that of lidocaine (0.18 mM), which is consistent with the weak anesthetic capacity of this analogue. In contrast, JMF2-1 was more potent than lidocaine in inhibiting allergen-induced histamine release and tracheal spasm. In in vivo settings methacholine-induced increase in lung resistance (145%) significantly reduced to 72% and 47% after lidocaine and JMF2-1 treatment, respectively. Both treatments inhibited by about 81% allergen-evoked eosinophil accumulation into the lung tissue. CONCLUSION: Replacement of the 2,6-dimethyl radicals by the 2-trifluormethyl group on the benzene ring of lidocaine significantly reduces anesthetic activity, preserving its ability to prevent key aspects of the allergic inflammatory response in the lung. CLINICAL IMPLICATIONS: Nebulized JMF2-1 might be a means of achieving the antiasthmatic effects of lidocaine without the anesthetic effects.

Avaliação do potencial imunorregulatório do análogo da lidocaína JMF2-1 em um modelo murino de asma / Evaluation of the immunoregulatory potential of the analogous one of lidocaine JMF2-1 in a murino model of asthma

A asma é caracterizada por uma inflamação crônica nas vias aéreas pulmonares causada por fatores ambientais em indivíduos geneticamente pré-dispostos. Estudos anteriores demonstraram que a aerolização com o análogo estrutural da lidocaína JMF2-1 poderia ser uma forma de atingir os efeitos antiasmáticos da lidocaína, sem apresentar a sua ação anestésico local. Nossos dados prévios apontam para um importante efeito antiinflamatório do JMF2-1 em modelos experimentais de asma, mas o modo de ação desta substância ainda não está elucidado. Nesse trabalho examinamos os efeitos imunorregulatórios do JMF2-1, em comparação com a lidocaína e a dexametasona, em um modelo murino de asma. Camundongos BALB/c sensibilizados e desafiados com ovoalbumina foram tratados com lidocaína e JMF2-1 aerosol, concomitante ao desafio e oito horas após cada desafio. O tratamento intraperitoneal com dexametasona foi realizado uma vez ao dia uma hora antes de cada desafio. Todos os tratamentos ocorreram dos dias 19 ao 21 pós-sensibilização, todas as análises ocorreram 24 horas após o último desafio. Primeiramente, observamos que a inalação da lidocaína e do JMF2-1 reduziu o número de eosinófilos nos pulmões dos animais desafiados com ovoalbumina. Além disso, ambos os fármacos inibiram a hiperreatividade brônquica dos animais desafiados. Os tratamentos in vivo com lidocaína e JMF2-1, assim como com dexametasona, inibiram a secreção de IL-4, IL-5 e IL-13 em culturas de explante pulmonar, obtidos dos camundongos sensibilizados e desafiados com ovoalbumina. Após um segundo desafio com ovoalbumina, dessa vez in vitro, a produção de IL-5 e IL-13 foi inibida quando os explantes eram provenientes de camundongos tratados com dexametasona, mas não lidocaína e JMF2-1. Células obtidas dos linfonodos de camundongos DO11.10 - TCR transgênico foram estimuladas com ovoalbumina e tratadas in vitro com JMF2-1, lidocaína ou dexametasona. A produção de citocinas e proliferação das células dos...