ABSTRACT
Inhibitors of sodium/glucose cotransporter 2 (SGLT2), such as empagliflozin and canagliflozin, have been widely used to block glucose reabsorption in the proximal tubules of kidneys in patients with diabetes. A meta-analysis suggested that SGLT2 inhibitors are associated with a decreased risk of asthma development. Therefore, we investigated whether SGLT2 inhibitors could suppress allergic asthma. Empagliflozin and canagliflozin suppressed the in vitro degranulation reaction induced by antigens in a concentration-dependent manner in RBL-2H3 mast cells. Empagliflozin and canagliflozin were administered to BALB/c mice sensitized to ovalbumin (OVA). The administration of empagliflozin or canagliflozin significantly suppressed OVA-induced airway hyper-responsiveness and increased the number of immune cells and pro-inflammatory cytokine mRNA expression levels in bronchoalveolar lavage fluid. The administration of empagliflozin and canagliflozin also suppressed OVA-induced histopathological changes in the lungs. Empagliflozin and canagliflozin also suppressed serum IgE levels. These results suggested that empagliflozin and canagliflozin may be applicable for the treatment of allergic asthma by suppressing immune responses.
Subject(s)
Asthma , Benzhydryl Compounds , Canagliflozin , Glucosides , Mice, Inbred BALB C , Ovalbumin , Sodium-Glucose Transporter 2 Inhibitors , Animals , Canagliflozin/pharmacology , Canagliflozin/therapeutic use , Sodium-Glucose Transporter 2 Inhibitors/pharmacology , Sodium-Glucose Transporter 2 Inhibitors/therapeutic use , Asthma/drug therapy , Asthma/immunology , Glucosides/pharmacology , Glucosides/therapeutic use , Benzhydryl Compounds/pharmacology , Benzhydryl Compounds/therapeutic use , Mice , Ovalbumin/adverse effects , Lung/pathology , Lung/drug effects , Lung/metabolism , Mast Cells/drug effects , Mast Cells/metabolism , Mast Cells/immunology , Immunoglobulin E/blood , Immunoglobulin E/immunology , Bronchoalveolar Lavage Fluid , Rats , Cytokines/metabolism , Cell Degranulation/drug effects , Disease Models, Animal , Sodium-Glucose Transporter 2/metabolism , MaleABSTRACT
Asthma is a chronic immunological disease related to oxidative stress and chronic inflammation; both processes promote airway remodeling with collagen deposition and matrix thickening, causing pulmonary damage and lost function. This study investigates the immunomodulation of C-phycocyanin (CPC), a natural blue pigment purified from cyanobacteria, as a potential alternative treatment to prevent the remodeling process against asthma. We conducted experiments using ovalbumin (OVA) to induce asthma in Sprague Dawley rats. Animals were divided into five groups: (1) sham + vehicle, (2) sham + CPC, (3) asthma + vehicle, (4) asthma + CPC, and (5) asthma + methylprednisolone (MP). Our findings reveal that asthma promotes hypoxemia, leukocytosis, and pulmonary myeloperoxidase (MPO) activity by increasing lipid peroxidation, reactive oxygen and nitrogen species, inflammation associated with Th2 response, and airway remodeling in the lungs. CPC and MP treatment partially prevented these physiological processes with similar action on the biomarkers evaluated. In conclusion, CPC treatment enhanced the antioxidant defense system, thereby preventing oxidative stress and reducing airway inflammation by regulating pro-inflammatory and anti-inflammatory cytokines, consequently avoiding asthma-induced airway remodeling.
Subject(s)
Airway Remodeling , Asthma , Disease Models, Animal , Ovalbumin , Oxidative Stress , Phycocyanin , Rats, Sprague-Dawley , Animals , Phycocyanin/pharmacology , Phycocyanin/therapeutic use , Asthma/drug therapy , Asthma/metabolism , Asthma/chemically induced , Oxidative Stress/drug effects , Ovalbumin/adverse effects , Rats , Airway Remodeling/drug effects , Inflammation/metabolism , Inflammation/drug therapy , Male , Lung/drug effects , Lung/pathology , Lung/metabolism , Cytokines/metabolismABSTRACT
BACKGROUND: Allergic asthma is an important respiratory system problem characterized by airway inflammation, breathlessness, and bronchoconstriction. Allergic asthma and its outcomes are triggered by type 2 allergic immune responses. Tectorigenin is a methoxy-isoflavone with anti-inflammatory effects. In this study, we investigated the effects of tectorigenin on the pathophysiology of allergic asthma in an animal model. METHODS: Asthmatic mice were treated with tectorigenin. Then airway hyperresponsiveness (AHR), eosinophil percentage, levels of interleukin (IL)-33, IL-25, IL-13, IL-5, IL-4, total and ovalbumin (OVA)-specific immunoglobulin (Ig)E, and lung histopathology were evaluated. RESULT: Tectorigenin significantly (P ã 0.05) reduced eosinophil infiltration (41 ± 7%) in the broncho-alveolar lavage fluid (BALF), serum IL-5 level (41 ± 5, pg/mL), and bronchial and vascular inflammation (scores of 1.3 ± 0.2 and 1.1 ± 0.3, respectively) but had no significant effects on AHR, serum levels of IL-33, -25, -13, and -4 (403 ± 24, 56 ± 7, 154 ± 11, and 89 ± 6 pg/mL, respectively), total and OVA-specific IgE (2684 ± 265 and 264 ± 19 ng/mL, respectively), goblet cell hyperplasia, and mucus production. CONCLUSION: Tectorigenin could control inflammation and the secretion of inflammatory mediators of asthma, so it can be regarded as a potential antiasthma treatment with the ability to control eosinophilia-related problems.
Subject(s)
Anti-Inflammatory Agents , Antioxidants , Asthma , Disease Models, Animal , Isoflavones , Mice, Inbred BALB C , Ovalbumin , Animals , Asthma/drug therapy , Asthma/chemically induced , Asthma/metabolism , Asthma/immunology , Asthma/pathology , Mice , Ovalbumin/toxicity , Ovalbumin/adverse effects , Isoflavones/pharmacology , Isoflavones/therapeutic use , Antioxidants/pharmacology , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Immunoglobulin E/blood , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Female , Lung/pathology , Lung/drug effects , Lung/metabolism , Lung/immunology , Cytokines/metabolismABSTRACT
Asthma is recognized as a chronic respiratory illness characterized by airway inflammation and airway hyperresponsiveness. Wogonoside, a flavonoid glycoside, is reported to significantly alleviate the inflammation response and oxidative stress. Herein, this study aimed to investigate the therapeutic effect and underlying mechanism of wogonoside on airway inflammation and mucus hypersecretion in a murine asthma model and in human bronchial epithelial cells (16HBE). BALB/c mice were sensitized and challenged with ovalbumin (OVA). Pulmonary function and the number of cells in the bronchoalveolar lavage fluid (BALF) were examined. Pathological changes in lung tissue in each group were evaluated via hematoxylin and eosin and periodic acid-Schiff staining, and changes in levels of cytokines in BALF and of immunoglobulin E in serum were determined via an enzyme-linked immunosorbent assay. The expression of relevant genes in lung tissue was analyzed via real-time PCR. Western blotting and immunofluorescence were employed to detect the expression of relevant proteins in lung tissue and 16HBE cells. Treatment with 10 and 20 mg/kg wogonoside significantly attenuated the OVA-induced increase of inflammatory cell infiltration, mucus secretion, and goblet cell percentage and improved pulmonary function. Wogonoside treatment reduced the level of T-helper 2 cytokines including interleukin (IL)-4, IL-5, and IL-13 in BALF and of IgE in serum and decreased the mRNA levels of cytokines (IL-4, IL-5, IL-6, IL-13, and IL-1ß and tumor necrosis factor-α), chemokines (CCL-2, CCL-11, and CCL-24), and mucoproteins (MUC5AC, MUC5B, and GOB5) in lung tissues. The expression of MUC5AC and the phosphorylation of STAT6 and NF-κB p65 in lung tissues and 16HBE cells were significantly downregulated after wogonoside treatment. Thus, wogonoside treatment may effectively decrease airway inflammation, airway remodeling, and mucus hypersecretion via blocking NF-κB/STAT6 activation.
Subject(s)
Asthma , Flavanones , Glucosides , NF-kappa B , Humans , Animals , Mice , NF-kappa B/metabolism , Ovalbumin/adverse effects , Ovalbumin/metabolism , Interleukin-13 , Interleukin-5/metabolism , Interleukin-5/pharmacology , Interleukin-5/therapeutic use , Asthma/chemically induced , Asthma/drug therapy , Asthma/genetics , Lung/metabolism , Inflammation/metabolism , Mucus/metabolism , Cytokines/genetics , Cytokines/metabolism , Bronchoalveolar Lavage Fluid , Mice, Inbred BALB C , Disease Models, Animal , STAT6 Transcription Factor/genetics , STAT6 Transcription Factor/metabolism , STAT6 Transcription Factor/pharmacologyABSTRACT
Rhizoma Dioscoreae Nipponicae (RDN) is a traditional Chinese medicine that widely applied in the treatment of human diseases. This study aims to explore the therapeutic potential of RDN in asthma and the underlying mechanisms. A mouse model of asthma was established by the stimulation of ovalbumin (OVA). HE staining was performed to detect the pathological injuries of tracheal tissues. The protein expression of collagen I, FN1, α-SMA (airway remodeling markers), and p-p38 (a marker of the p38 MAPK pathway) were detected by Western blot. Eosinophils were then isolated from the model mice. Cell viability and ROS level were measured by CCK-8 and Flow cytometry, respectively. The mRNA expression of GPX4 and ACSL4 (ferroptosis markers) in eosinophils were measured by qRT-PCR. RDN significantly reduced the numbers of total cells and eosnophils in bronchoalveolar lavage fluid (BALF), inhibited inflammatory cell infiltration, and down-regulated remodeling markers (Collagen I, FN1, and α-SMA) in OVA-induced mice. The p38 MAPK pathway was blocked by the intervention of RDN in the model mice, and its blocking weakens the poor manifestations of OVA-induced asthma. In addition, RDN induced the ferroptosis of eosnophils both in vitro and in vivo. Blocking of the p38 MAPK pathway also enhanced the ferroptosis of eosnophils in vitro, evidenced by the decreased cell viability and GPX4 expression, and increased ROS level and ACSL4 expression. RDN induced the ferroptosis of eosinophils through inhibiting the p38 MAPK pathway, contributing to the remission of asthma.
Subject(s)
Asthma , Ferroptosis , Animals , Humans , Mice , Asthma/metabolism , Collagen/metabolism , Disease Models, Animal , Eosinophils/metabolism , Lung/pathology , Ovalbumin/adverse effects , p38 Mitogen-Activated Protein Kinases/metabolism , Reactive Oxygen Species/metabolism , Signal TransductionABSTRACT
BACKGROUND: Food allergy has become a global public health problem. This study aimed to explore the possible anti-allergic effect of vitamin C (VC). A rat basophilic leukemia (RBL)-2H3 cell degranulation model was used to assess the effect of VC on degranulation in vitro, and an ovalbumin (OVA)-induced BALB/c mouse allergy model was used to assess the anti-allergy effect of VC in vivo. RESULTS: In vitro, VC significantly attenuated the release of ß-hexosaminidase, tryptase and histamine, and also reduced cytokine production (interleukins 4 and 6, tumor necrosis factor α) significantly (P < 0.05), with the inhibitory effect demonstrating a positive correlation with VC dose. In vivo, compared with the OVA group, the levels of serum immunoglobulins E and G1 of the VC low-dose (VCL) group (50 mg kg-1) and high-dose (VCH) group (200 mg·kg-1) were significantly reduced (P < 0.05). Furthermore, the plasma histamine level was also significantly decreased (P < 0.05). Moreover, TH2 cell polarization in mice of the VCL and VCH groups was significantly inhibited (P < 0.05), promoting the TH1/TH2 cell polarization balance. Additionally, VC treatment enhanced the expression of CD80 (P < 0.05) in spleen and small intestine tissues, while significantly inhibiting the expression of CD86 (P < 0.05); notably, high-dose VC treatment was more effective. CONCLUSION: VC exerted an anti-allergic effect through inhibiting degranulation and regulating TH1/TH2 cell polarization balance. © 2024 Society of Chemical Industry.
Subject(s)
Anti-Allergic Agents , Ascorbic Acid , Cell Degranulation , Food Hypersensitivity , Mice, Inbred BALB C , Th1 Cells , Th2 Cells , Animals , Th2 Cells/immunology , Th2 Cells/drug effects , Anti-Allergic Agents/pharmacology , Mice , Ascorbic Acid/pharmacology , Cell Degranulation/drug effects , Th1 Cells/immunology , Th1 Cells/drug effects , Rats , Food Hypersensitivity/drug therapy , Food Hypersensitivity/immunology , Immunoglobulin E/immunology , Humans , Female , Male , Ovalbumin/immunology , Ovalbumin/adverse effects , Cytokines/metabolism , Cytokines/immunology , beta-N-Acetylhexosaminidases/metabolismABSTRACT
OBJECTIVES: Desmodium triquetrum DC (Fabaceae) is a plant commonly used in Indian traditional medicine to treat allergies. Asthma is a severe condition, with an estimated 300 million deaths annually, which could increase to 400 million by 2025. Flavonoids, a class of compounds found in many plants, have been found to have beneficial effects in treating asthma. In this study, researchers focused on three flavonoids, Baicalein, Naringin, and Neohesperidin, derived from Desmodium triquetrum DC, to investigate their potential as a treatment for asthma. METHODS: The study used an aerosolized ovalbumin-induced asthma model to evaluate the effects of the flavonoids on various substances in bronchoalveolar lavage fluid, including total differential leukocyte, nitrite, nitrate, TNF, IL-4, and IL-13. The researchers also measured the levels of myeloperoxidase and malondialdehyde in the lungs. RESULTS: The results showed that ovalbumin-induced airway hyper-responsiveness led to a significant increase in pro-inflammatory cytokine levels. However, the flavonoids significantly decreased the severity of airway inflammation. Histopathology results also supported the effectiveness of the flavonoids. These findings suggest that these flavonoids could be a supplementary and alternative treatment for asthma by inhibiting the pro-inflammatory pathway. CONCLUSIONS: The findings suggest that the isolated compounds have the potential to act cumulatively to decrease the levels of the tested cytokines, normalize eosinophil and activated lymphocyte counts, and significantly reduce MPO and MDA. This indicates a possible respiratory mechanism of action for the drugs.
Subject(s)
Asthma , Flavonoids , Animals , Mice , Ovalbumin/adverse effects , Ovalbumin/metabolism , Flavonoids/pharmacology , Flavonoids/therapeutic use , Asthma/chemically induced , Asthma/drug therapy , Lung/metabolism , Lung/pathology , Cytokines , Inflammation/drug therapy , Mice, Inbred BALB C , Disease Models, AnimalABSTRACT
BACKGROUND: Growing up on traditional farms protects children from the development of asthma and allergies. However, we have identified distinct asthma-protective factors, such as poultry exposure. This study aims to examine the biological effect of rural exposure in China. METHODS: We recruited 67 rural children (7.4 ± 0.9 years) and 79 urban children (6.8 ± 0.6 years). Depending on the personal history of exposure to domestic poultry (DP), rural children were further divided into those with DP exposure (DP+ , n = 30) and those without (DP- , n = 37). Blood samples were collected to assess differential cell counts and expression of immune-related genes. Dust samples were collected from poultry stables inside rural households. In vivo activities of nasal administration of DP dust extracts were tested in an ovalbumin-induced asthma model. RESULTS: There was a stepwise increase in the percentage of eosinophils (%) from rural DP+ children (median = 1.65, IQR = [1.28, 3.75]) to rural DP- children (3.40, [1.70, 6.50]; DP+ vs. DP- , p = .087) and to the highest of their urban counterparts (4.00, [2.00, 7.25]; urban vs. DP+ , p = .017). Similarly, rural children exhibited reduced mRNA expression of immune markers, both at baseline and following lipopolysaccharide (LPS) stimulation. Whereas LPS stimulation induced increased secretion of Th1 and proinflammatory cytokines in rural DP+ children compared to rural DP- children and urban children. Bronchoalveolar lavage of mice with intranasal instillation of dust extracts from DP household showed a significant decrease in eosinophils as compared to those of control mice (p < .05). Furthermore, DP dust strongly inhibited gene expression of Th2 signature cytokines and induced IL-17 expression in the murine asthma model. CONCLUSIONS: Immune responses of rural children were dampened compared to urban children and those exposed to DP had further downregulated immune responsiveness. DP dust extracts ameliorated Th2-driven allergic airway inflammation in mice. Determining active protective components in the rural environment may provide directions for the development of primary prevention of asthma.
Subject(s)
Asthma , Hypersensitivity , Child , Humans , Animals , Mice , Lipopolysaccharides/adverse effects , Allergens , Cytokines/metabolism , Dust , Inflammation , Disease Models, Animal , Immunity , Mice, Inbred BALB C , Ovalbumin/adverse effectsABSTRACT
Asthma is a chronic pulmonary disease with marked infiltrating inflammatory cells and reduced respiratory performance. Echinochrome (Ech) is a dark-red pigment isolated from the sea urchin spines, shells, and ova. It has antioxidant, antimicrobial, and anti-inflammatory properties, but whether it can be used in asthma treatment has yet to be investigated. In this research, we aimed to study the inhibitory actions of Ech on allergic asthma symptoms in mice. Mice were divided into 4 groups (n = 8 for each): control, ovalbumin-challenged, and Ech-treated (0.1 and 1 mg/kg). At the end of the experiment, nasal scratching, lung oxidative stress, airway inflammation, and remodeling were assessed. In ovalbumin-challenged BALB/C mice, treatment with Ech significantly decreased nasal scratching, lung oxidative stress, inflammatory cell infiltration, mucus hyperproduction and hyperplasia of goblet cells, IgE levels, and inflammatory cytokines. It also inhibited NF-κB phosphorylation. This is the first study to investigate the immunomodulatory effect of Ech against allergic asthma in mice. According to our findings, we imply that Ech may be utilized as a treatment for allergic asthma.
Subject(s)
Anti-Asthmatic Agents , Asthma , Animals , Mice , Anti-Asthmatic Agents/therapeutic use , Ovalbumin/adverse effects , Mice, Inbred BALB C , Immunoglobulin E , Asthma/chemically induced , Asthma/drug therapy , Asthma/pathology , Inflammation/pathology , Cytokines/metabolism , Oxidative Stress , Disease Models, Animal , Bronchoalveolar Lavage FluidABSTRACT
INTRODUCTION: For centuries, Salvia rosmarinus Spenn has been applied as folk medicine to cure different diseases due to its anti-inflammatory, antibacterial, antioxidant, antifungal, and antitumor effects. To find bioactive medicinal herbs exerting a protective effect on airway inflammation and remodeling, we assessed the anti-oxidative and anti-inflammatory effects of an aqueous spray-dried extract of Salvia rosmarinus Spenn. (rosemary) in an ovalbumin-induced asthmatic rat model. METHODS: Rats were randomly divided into normal control (control), asthma, asthma+rosemary extract (RE) (13 mg/kg), asthma+RE (50 mg/kg), and asthma+budesonide groups. After 50 days, animals were anesthetized, and then blood, bronchoalveolar lavage fluid (BALF), and lung tissues were collected for subsequent serological and pathological studies. Histopathology of lung tissues was evaluated by H&E staining. The oxidative stress parameters and airway inflammation factors in BALF and lung tissue were explored. RESULTS: Using thin layer chromatography, the presence of rosmarinic acid was confirmed in aqueous extract of rosemary. Furthermore, RE markedly decreased immunoglobulin E levels (50 mg/kg; p < 0.001 vs. asthma group) and inflammatory cytokines (50 mg/kg; p < 0.001 vs. asthma group) and increased antioxidant enzymes (50 mg/kg, p < 0.001 vs. asthma group). Furthermore, RE at a concentration of 50 mg/kg obviously reduced the number of inflammatory cells, goblet cells, and pathological changes compared to the asthma group. CONCLUSION: The results showed that RE administration might prevent or alleviate allergic asthma-related pathological change, probably via antioxidant and anti-inflammatory mechanisms.
Subject(s)
Asthma , Rosmarinus , Salvia , Rats , Animals , Mice , Antioxidants/pharmacology , Antioxidants/therapeutic use , Asthma/chemically induced , Asthma/drug therapy , Lung/pathology , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Inflammation/pathology , Bronchoalveolar Lavage Fluid , Oxidative Stress , Ovalbumin/adverse effects , Disease Models, Animal , Mice, Inbred BALB CABSTRACT
INTRODUCTION: Asthma has been attributed to Th1/Th2 imbalance and inappropriate Th2 responses to environmental allergens. MicroRNAs (miRNAs), 21 to 23 RNA molecules, are first found in mammals and have been implicated in various biological activities. Our previous study found that miR-410 effectively ameliorates airway inflammation in the ovalbumin (OVA)-induced asthma murine model. However, the role of miR-410 in regulating helper T (Th) cell differentiation is not clear. In the present study, we aimed to explore the regulatory effects of miR-410 on the differentiation of Th cells through both in vivo and in vitro studies. METHODS: Dual-luciferase reporter assay was used to find if miR-410 has any direct binding position with VEGF mRNAs. PBMC and CD4+ T cells were isolated and stimulated with OVA. The miR-410 mimics and inhibitors were transfected into CD4+ T cells. The differentiation of Th cells was evaluated by enzyme-linked immunosorbent assay (ELISA) for the concentration of IL-4, IFN-γ, and TGF-ß levels in supernatants. Western Blot was used to detect protein expression and phosphorylation of PI3K and AKT. BALB/c mice were kept in a specific pathogen-free condition and received sterile OVA-free food and water. OVA-induced asthmatic mice model was established. ELISA was used to measure the bronchoalveolar lavage fluid (BALF) concentrations of IL-4, IFN-γ, TGF-ß, and VEGF. Hematoxylin and eosin staining and immunohistochemical staining were conducted to analyze inflammatory cell infiltration, pathological changes, and the expression of VEGF. RESULTS: Dual-luciferase reporter assay showed that miR-410 has no direct binding position with VEGF mRNAs. In the OVA-primed mononuclear cells compared to normal cells, IFN-γ and TGF-ß were decreased while IL-4 and VEGF were increased. This change was reversed while miRNA-410 mimics were transfected into CD4+ T cells. Besides, the OVA-primed CD4+ T cells treated with miR-410 decrease the proliferation of cytokine of Th2 cells as well as phosphorylation of PI3K, and AKT. In OVA-induced asthma mice, IFN-γ and TGF-ß were decreased in BALF while the IL-4 and VEGF were increased. OVA-induced mice with asthma treated with miR-410 mimics showed marked reductions in the infiltration of inflammatory cells as well as IL-4 and VEGF in BALF. The immunohistochemical staining of the expression of VEGF also decreased in OVA-induced asthma mice with the instillation of miR-410. CONCLUSIONS: In this study, we revealed that miR-410 could regulate the differentiation of Th cells via the PI3K-AKT-VEGF signaling pathway in asthma.
Subject(s)
Asthma , MicroRNAs , Animals , Mice , Asthma/metabolism , Bronchoalveolar Lavage Fluid , Cell Differentiation , Disease Models, Animal , Interleukin-4 , Leukocytes, Mononuclear , Luciferases/metabolism , Lung/pathology , Mammals/genetics , Mammals/metabolism , Mice, Inbred BALB C , MicroRNAs/genetics , Ovalbumin/adverse effects , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Signal Transduction , Transforming Growth Factor beta , Vascular Endothelial Growth Factor A/adverse effectsABSTRACT
Asthma is an inflammatory disease characterized by chronic inflammation in lung tissues and excessive mucus production. High-fat diets have long been assumed to be a potential risk factor for asthma. However, to date, very few direct evidence indicating the involvement of high sucrose intake (HSI) in asthma progression exists. In this study, we investigate the effect of HSI on ovalbumin (OVA)-sensitized allergic asthma mice. We observed that HSI increased the expression of inflammatory genes (IL-1ß, IL-6, TNF-α) in adipose tissues and led to reactive oxygen species generation in the liver and lung. In addition, HSI accelerated the TLR4/NF-κB signaling pathway leading to MMP9 activation, which promotes the chemokines and TGF-ß secretion in the lungs of OVA-sensitized allergic asthma mice. More importantly, HSI significantly promoted the pathogenic Th2 and Th17 responses. The increase of IL-17A secretion by HSI increased the expression of chemokines (MCP-1, CXCL1, CXCL5, CXCL8). It resulted in eosinophil and mast cell infiltration in the lung and trachea. We also demonstrated that HSI increased mucus hypersecretion, which was validated by increased main mucin protein (MUC5AC) secreted in the lungs. Our findings suggest that HSI exacerbates the development of Th2/Th17-predominant asthma by upregulating the TLR4-mediated NF-κB pathway, leading to excessive MMP9 production.
Subject(s)
Asthma , Matrix Metalloproteinase 9 , Mice , Animals , Ovalbumin/adverse effects , Matrix Metalloproteinase 9/metabolism , NF-kappa B/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Mice, Inbred C57BL , Asthma/metabolism , Lung , Inflammation/metabolism , Chemokines/metabolism , Sucrose/adverse effects , Mice, Inbred BALB C , Disease Models, Animal , Bronchoalveolar Lavage FluidABSTRACT
BACKGROUND: Transforming growth factor-ß (Tgf-ß) plays an important role in the pathogenesis of asthma through the regulation of T cells and airway epithelium. Its functions in alveolar macrophage (AM) during allergic airway inflammation remain unknown. METHODS: A murine asthma model was induced with ovalbumin (ova) in TßRICA/Fsp1-Cre transgenic mice expressing constitutively active Tgf-ß receptor type I (TßRICA) under the control of Fsp1-Cre transgene. Cells in the bronchoalveolar lavage (BAL) were collected to study immune cell infiltration in the lungs. Cytokine levels in BAL fluid were measured by enzyme-linked immunoassay (ELISA). Lungs were sectioned and stained with hematoxylin and eosin, periodic acid-Schiff, and trichrome for histopathologic evaluation. AMs were assessed by flow cytometry and were sorted for quantitative polymerase chain reaction analysis. RESULTS: Our data indicated that TßRICA transcripts were induced in AMs of TßRICA/Fsp1-Cre mice. Following the ova challenges, TßRICA/Fsp1-Cre mice exhibited reduced cellular infiltration of the airway, reduced pulmonary fibrosis, and reduced bronchial mucus secretion as compared to ova-challenged wild-type mice. An alternatively activated macrophage (M2) polarization was significantly elevated in the lungs of ova-challenged TßRICA/Fsp1-Cre mice as reflected by increased numbers of AMs expressing M2 subtype marker, CD163, in the lungs and enhanced expression of CCR2 and CD206 in AMs. Moreover, TßRICA/Fsp1-Cre AMs showed augmented expression of transcription factors, Foxo1, and IRF4, which are known to be positive regulators for M2 polarization. CONCLUSIONS: Expression of TßRICA in AMs promoted M2 polarization and ameliorated allergic airway inflammation in an ova-induced asthma mouse model.
Subject(s)
Asthma , Macrophages, Alveolar , Mice , Humans , Animals , Ovalbumin/adverse effects , Asthma/metabolism , Lung/pathology , Inflammation , Bronchoalveolar Lavage Fluid , Disease Models, Animal , Mice, Inbred BALB CABSTRACT
Asthma is a common respiratory disease associated with airway inflammation. Nerolidol is an acyclic sesquiterpenoid with anti-inflammatory properties. BALB/C mice were sensitized with ovalbumin (OVA) to induce asthma symptoms and given different doses of Nerolidol. We found that Nerolidol reduced OVA-induced inflammatory cell infiltration, the number of goblet cells and collagen deposition in lung tissue. Nerolidol reduced the OVA-specific IgE levels in serum and alveolar lavage fluid in an asthma model. Immunohistochemical staining of α-SMA (the marker of airway smooth muscle) showed that Nerolidol caused bronchial basement membrane thinning in asthmatic mice. The hyperplasia of airway smooth muscle cells (ASMCs) is an important feature of airway remodeling in asthma. ASMCs were treated with 10 ng/mL TGF-ß to simulate the pathological environment of asthma in vitro and then treated with different doses of Nerolidol. Nerolidol inhibited the activity of TGF-ß/Smad signaling pathway both in the lung tissue of OVA-induced mouse and TGF-ß-stimulated ASMCs. 16s rRNA sequencing was performed on feces of normal mice, the changes of intestinal flora in OVA-induced asthmatic mice and Nerolidol-treated asthmatic mice were studied. The results showed that Nerolidol reversed the reduced gut microbial alpha diversity in asthmatic mice. Nerolidol changed the relative abundance of gut bacteria at different taxonomic levels. At the phylum level, the dominant bacteria were Bacteroidota, Firmicutes, and Proteobacteria. At the genus level, the dominant bacteria were Lactobacillus, Muribaculaceae, Bacteroides, and Lachnospiraceae. We conclude that Nerolidol attenuates OVA-induced airway inflammation and alters gut microbes in mice with asthma via TGF-ß/Smad signaling.
Subject(s)
Asthma , Gastrointestinal Microbiome , Sesquiterpenes , Animals , Mice , Ovalbumin/adverse effects , Ovalbumin/metabolism , Airway Remodeling , RNA, Ribosomal, 16S/metabolism , Mice, Inbred BALB C , Asthma/chemically induced , Asthma/drug therapy , Asthma/metabolism , Lung/metabolism , Lung/pathology , Sesquiterpenes/pharmacology , Inflammation/chemically induced , Inflammation/drug therapy , Inflammation/metabolism , Bronchoalveolar Lavage Fluid/chemistry , Transforming Growth Factor beta/metabolism , Disease Models, AnimalABSTRACT
Purpose: To investigate the in vivo efficacy of epinastine cream in type I allergic models. Methods: The dose, timing, and antiallergic effect of epinastine cream on the conjunctiva were evaluated postapplication to the eyelid skin of guinea pigs with histamine- or ovalbumin-induced allergic conjunctivitis. Additionally, we assessed its antiallergic effects on the skin postapplication to the dorsal skin of guinea pigs with ovalbumin-induced passive cutaneous anaphylaxis. Efficacy was estimated by determining the amount of dye that leaked from conjunctival or dorsal skin tissue vessels as a measure of vascular permeability, scoring the severity of allergic symptoms, and observing the scratching behaviors using clinical parameters. Results: In the histamine-induced conjunctivitis model, epinastine cream strongly inhibited conjunctival vascular permeability in a dose-dependent manner. The inhibitory effect of 0.5% epinastine cream 24 h postapplication was significantly higher than that of 0.1% epinastine hydrochloride ophthalmic solution 8 h postadministration. Additionally, the 0.5% epinastine cream inhibited conjunctival vascular permeability 15 min postapplication, and the effect was sustained over 24 h. Furthermore, the 0.5% epinastine cream effectively suppressed clinical symptom scores and exhibited ameliorated scratching bouts in conjunctival allergic reactions in the experimental allergic conjunctivitis model. Additionally, it significantly inhibited vascular permeability in skin allergic reactions in the passive cutaneous anaphylaxis model. Conclusions: The results suggest that epinastine cream is a strong, long-lasting, and skin-penetrating inhibitor of type I allergic reactions. The 0.5% epinastine cream applied once daily could be a promising, potent, and long-acting therapeutic agent for allergic conjunctivitis.
Subject(s)
Anti-Allergic Agents , Conjunctivitis, Allergic , Dibenzazepines , Imidazoles , Animals , Guinea Pigs , Conjunctivitis, Allergic/drug therapy , Conjunctivitis, Allergic/chemically induced , Conjunctivitis, Allergic/diagnosis , Histamine/adverse effects , Histamine H1 Antagonists/adverse effects , Ovalbumin/adverse effects , Anti-Allergic Agents/pharmacology , Anti-Allergic Agents/therapeutic useABSTRACT
BACKGROUND: Angiotensin (Ang)-(1-7) can reduce airway inflammation and airway remodeling in allergic asthma. Autophagy-related 5 (ATG5) has attracted wide attentions in asthma. However, the effects of Ang-(1-7) on ATG5-mediated autophagy in allergic asthma are unclear. METHODS: In this study, human bronchial epithelial cell (BEAS-2B) and human bronchial smooth muscle cell (HBSMC) were treated with different dose of Ang-(1-7) to observe changes of cell viability. Changes of ATG5 protein expression were measured in 10 ng/mL of interleukin (IL)-13-treated cells. Transfection of ATG5 small interference RNA (siRNA) or ATG5 cDNA in cells was used to analyze the effects of ATG5 on secretion of cytokines in the IL-13-treated cells. The effects of Ang-(1-7) were compared to the effects of ATG5 siRNA transfection or ATG5 cDNA transfection in the IL-13-treated cells. In wild-type (WT) mice and ATG5 knockout (ATG5-/-) mice, ovalbumin (OVA)-induced airway inflammation, fibrosis and autophagy were observed. In the OVA-induced WT mice, Ang-(1-7) treatment was performed to observe its effects on airway inflammation, fibrosis and autophagy. RESULTS: The results showed that ATG5 protein level was decreased with Ang-(1-7) dose administration in the IL-13-treated BEAS-2B and IL13-treated HBSMC. Ang-(1-7) played similar results to ATG5 siRNA that it suppressed the secretion of IL-25 and IL-13 in the IL-13-treated BEAS-2B cells, and inhibited the expression of transforming growth factor (TGF)-ß1 and α-smooth muscle actin (α-SMA) protein in the IL-13-treated HBSMC cells. ATG5 cDNA treatment significantly increased the secretion of IL-25 and IL-13 and expression of TGF-ß1 and α-SMA protein in IL-13-treated cells. Ang-(1-7) treatment suppressed the effects of ATG5 cDNA in the IL-13-treated cells. In OVA-induced WT mice, Ang-(1-7) treatment suppressed airway inflammation, remodeling and autophagy. ATG5 knockout also suppressed the airway inflammation, remodeling and autophagy. CONCLUSIONS: Ang-(1-7) treatment suppressed airway inflammation and remodeling in allergic asthma through inhibiting ATG5, providing an underlying mechanism of Ang-(1-7) for allergic asthma treatment.
Subject(s)
Asthma , Lung , Humans , Animals , Mice , Lung/pathology , Ovalbumin/adverse effects , Interleukin-13 , Airway Remodeling , Autophagy-Related Protein 5/genetics , Autophagy-Related Protein 5/pharmacology , Autophagy-Related Protein 5/therapeutic use , DNA, Complementary/adverse effects , Asthma/genetics , Transforming Growth Factor beta1/metabolism , Inflammation/drug therapy , RNA, Small Interfering/adverse effects , Fibrosis , Disease Models, Animal , Mice, Inbred BALB CABSTRACT
Interleukin-17A (IL-17A) levels are elevated in patients with asthma. Ferroptosis has been identified as the non-apoptotic cell death type associated with asthma. Data regarding the relation of ferroptosis with asthma and the effect of IL-17A on modulating ferroptosis in asthma remain largely unclear. The present work focused on investigating the role of IL-17A in allergic asthma-related ferroptosis and its associated molecular mechanisms using public datasets, clinical samples, human bronchial epithelial cells, and an allergic asthma mouse model. We found that IL-17A was significantly upregulated within serum in asthma cases. Adding IL-17A significantly increased ferroptosis within human bronchial epithelial cells (BEAS-2B). In ovalbumin (OVA)-induced allergic asthmatic mice, IL-17A regulated and activated lipid peroxidation induced ferroptosis, whereas IL-17A knockdown effectively inhibited ferroptosis in vivo by protection of airway epithelial cells via the xCT-GSH-GPX4 antioxidant system and reduced airway inflammation. Mouse mRNA sequencing results indicated that the tumor necrosis factor (TNF) pathway was the differential KEGG pathway in the OVA group compared to healthy controls and the OVA group compared to the IL-17A knockout OVA group. We further used N-acetylcysteine (TNF inhibitor) to inhibit the TNF signaling pathway, which was found to protect BEAS-2B cells from IL-17A induced lipid peroxidation and ferroptosis damage. Our findings reveal a novel mechanism for the suppression of ferroptosis in airway epithelial cells, which may represent a new strategy for the use of IL-17A inhibitors against allergic asthma.
Subject(s)
Asthma , Ferroptosis , Animals , Humans , Mice , Asthma/pathology , Disease Models, Animal , Epithelial Cells/metabolism , Inflammation/genetics , Interleukin-17/genetics , Interleukin-17/adverse effects , Interleukin-17/metabolism , Lung/metabolism , Mice, Inbred BALB C , Ovalbumin/adverse effectsABSTRACT
INTRODUCTION: Many asthmatic patients are exposed to cigarette smoke actively or passively, which contributes to asthma exacerbation and poor control. This study is to explore the effects of cigarette smoke on pathological changes in murine surrogate of asthma. METHODS: C57BL/6 mice were sensitised and challenged with ovalbumin (OVA) to establish a surrogate of asthma and then administered with cigarette smoke extract (CSE). Airways hyperresponsiveness (AHR) was measured using the Flexivent system. Histological staining (haematoxylin-eosin [HE], periodic acid Schiff [PAS], Congo red and Masson's trichrome) was employed to measure pathological changes in sections of lung tissue of experimental mice. Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of total and OVA-specific IgE, cytokines and chemokines (eotaxin-1, IL-13, IL-1ß, TNF-α, IL-17A, IL-33) in the lung tissue homogenates. Immunoreactivity for vWF and α-SMA in lung tissue sections was detected by immunohistochemistry. RESULTS: Exposure of the animals to CSE significantly reduced OVA-induced AHR, the number of eosinophils in bronchoalveolar lavage fluid (BALF) and eosinophils infiltrating into the lung tissue, as well as concentrations of some cytokines in lung homogenate. In contrast, it significantly enhanced the number of macrophages and M2 in BALF, as well as collagen deposition, smooth muscle thickness and alveolar destruction in lung tissue. CONCLUSION: CSE inhibits OVA-induced AHR, changes inflammation 'phenotypes', while accelerates some aspects of airways remodelling, which might contribute to worse symptoms and be refractory to anti-inflammation therapies for asthmatics.
Subject(s)
Asthma , Cigarette Smoking , Humans , Animals , Mice , Ovalbumin/adverse effects , Mice, Inbred C57BL , Lung , Inflammation , Cytokines , Bronchoalveolar Lavage Fluid , Phenotype , Disease Models, Animal , Mice, Inbred BALB CABSTRACT
Asthma, a chronic airway inflammatory disease, has a complicated pathogenesis and limited therapeutic treatment. Evidence shows that the intestinal microbiota exhibits crucial functional interaction with asthma syndrome. Liubao tea (LBT), a type of postfermented tea in China, positively modulates gut microbiota. However, the potential benefits of LBT extract (LBTE) for allergic asthma are still not understood. Herein, the anti-inflammatory effects of LBTE and its modulation of the gut microbiota of asthmatic mice induced by ovalbumin were explored. The results demonstrate that LBTE significantly inhibited airway hyper-responsiveness and restrained the proliferation of proinflammatory cytokines and inflammatory cells associated with allergic asthma. Additionally, LBTE suppressed inflammatory infiltration, mucus secretion, and excessive goblet cell production by downregulating the gene expression of inflammatory indicators. Interestingly, fecal microbiota transplantation results further implied that the modulation of LBTE on gut microbiota played an essential role in alleviating airway inflammatory symptoms of allergic asthma.
Subject(s)
Asthma , Gastrointestinal Microbiome , Animals , Mice , Ovalbumin/adverse effects , Asthma/metabolism , Cytokines/metabolism , Tea/metabolism , Mice, Inbred BALB C , Disease Models, Animal , Lung , Bronchoalveolar Lavage FluidABSTRACT
Allergic asthma is a chronic inflammatory disease of airways involving complex mechanisms, including MAS-related GPR family member X2 (MRGPRX2) and its orthologue MRGPRB2 on mast cells (MCs). Although miRNAs have been previously shown to related to allergic asthma, the role of miR-212/132 in this process has not been studied. In this study, the predicted pairing of miRNAs and MRGPRX2 (MRGPRB2) mRNAs was carried out by online databases and the function was verify using in vivo and in vitro experiments. Database prediction showed that miR-212/132 interact with MRGPRX2 and MRGPRB2. miR-212/132 mimics alleviated MRGPRB2 mRNA expression as well as pathology changes in lungs and AHR of mice with airway inflammation in vivo. The expression level of MRGPRB2 in the mice lungs after inhaled OVA was also decreased by miR-212/132 mimics. Meanwhile, miR-212/132 inhibited MCs degranulation and cytokines release triggered by C48/80 in vitro. Further, ASAP1 (ARF GTPase-Activating Protein 1) was selected from the junction related pathways using RNAseq and KEGG enrichment. ASAP1 mRNA level was upregulated in airway inflammation and MCs activation and decreased by miR-212/132 mimics. miR-212/132 attenuated OVA-induced airway inflammation by inhibiting MCs activation through MRGPRX2 and ASAP1.