Inhaled pan-phosphodiesterase inhibitors ameliorate ovalbumin-induced airway inflammation and remodeling in murine model of allergic asthma.

Asthma is a heterogeneous, chronic respiratory disease characterized by airway inflammation and remodeling. Phosphodiesterase (PDE) inhibitors represent one of the intensively studied groups of potential anti-asthmatic agents due to their affecting both airway inflammation and remodeling. However, the effect of inhaled pan-PDE inhibitors on allergen induced asthma has not been reported to date. In this study we investigated the impact of two, representative strong pan-PDE inhibitors from the group of 7,8-disubstituted derivatives of 1,3-dimethyl-3,7-dihydro-1H-purine-2,6-dione: compound 38 and 145, on airway inflammation and remodeling in murine model of ovalbumin (OVA)-challenged allergic asthma. Female Balb/c mice were sensitized and challenged with OVA, 38 and 145 were administrated by inhalation, before each OVA challenge. The inhaled pan-PDE inhibitors markedly reduced the OVA-induced airway inflammatory cell infiltration, eosinophil recruitment, Th2 cytokine level in bronchoalveolar lavage fluid, as well as both, total and OVA-specific IgE levels in plasma. In addition, inhaled 38 and 145 decreased many typical features of airway remodeling, including goblet cell metaplasia, mucus hypersecretion, collagen overproduction and deposition, as well as Tgfb1, VEGF, and α-SMA expression in airways of allergen challenged mice. We also demonstrated that both 38 and 145 alleviate airway inflammation and remodelling by inhibition of the TGF-β/Smad signaling pathway activated in OVA-challenged mice. Taken together, these results suggest that the investigated pan-PDE inhibitors administered by inhalation are dual acting agents targeting both airway inflammation and remodeling in OVA-challenged allergic asthma and may represent promising, anti-asthmatic drug candidates.

Wójcik-Pszczoła K ，Pociecha K ，Chłoń-Rzepa G ，Zadrożna M ，Nowak B ，Plutecka H ，Koczurkiewicz-Adamczyk P ，Przejczowska-Pomierny K ，Pękala E ，Gosens R ，Wyska E ... -  《-》

MFG-E8/integrin β3 signaling contributes to airway inflammation response and airway remodeling in an ovalbumin-induced murine model of asthma.

Asthma is the most common chronic childhood disease worldwide, characterized by airway remodeling and chronic inflammation, orchestrated primarily by Th2 cytokines. The aim of the current study was to explore the influences of milk fat globule epidermal growth factor 8 (MFG-E8)/integrin β3 signaling involved in airway inflammation and remodeling in asthma. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed by OVA nebulization. The levels of MFG-E8 expression were declined markedly in the OVA-induced allergy murine model. In addition, administration of MFG-E8 strongly reduced the accumulation of T-helper type 2 (Th2)-associated cytokines (such as interleukin-4, -5, and -13) as well as chemokine CCL11 (eotaxin) in bronchoalveolar lavage fluid and tissues in the OVA-sensitized mice. Moreover, MFG-E8 remarkably repressed the total immunoglobulin E and OVA-specific immunoglobulin E in serum in OVA-challenged mice. Meanwhile, treatment with recombinant murine MFG-E8 noticeably prevented inflammatory cell infiltration into the airways, as showed by a marked decrease in the numbers of total immune cells, eosinophils, neutrophils, macrophages, and lymphocytes in the bronchoalveolar lavage fluid in response to OVA challenge. Importantly, MFG-E8 apparently alleviated OVA-driven airway remodeling, which were evidenced by declined secretion of important mediators of airway remodeling, including transforming growth factor-β1, matrix metalloproteinase 9, ADAM8, and vascular endothelial growth factor, and reduced airway collagen deposition and inhibited goblet cell hyperplasia in OVA-induced asthma in mice. Mechanistically, integrin 3 contributes to the protective effect of MFG-E8 in inhibiting airway inflammation and remodeling in OVA-driven features of allergic asthma. Overall, MFG-E8, as a candidate molecule to evaluate airway inflammation and remodeling, could be a potential target for the management and prevention of asthma exacerbations, suggesting that MFG-E8/integrin β3 signaling may serve as a promising therapeutic agent for childhood asthma.

Zhi Y ，Huang H ，Liang L 《-》

YiQi GuBen formula alleviates airway inflammation and airway remodeling in OVA-induced asthma mice through TLR4/NF-κB signaling pathway.

Kong Y ，Wang Z ，Yu H ，Dong A ，Song Y ，Guo L ，Zhu J ，Sun L ，Guo Y ... -  《-》

Protective effects and active ingredients of Salvia miltiorrhiza Bunge extracts on airway responsiveness, inflammation and remodeling in mice with ovalbumin-induced allergic asthma.

Salvia miltiorrhiza Bunge (S. miltiorrhiza), a traditional Chinese medicine, has demonstrated antioxidant, anti-inflammatory, and antibacterial activities. However, its effects against asthma that shows chronic inflammation and oxidative damage remain unknown. To assess the effects of S. miltiorrhiza extracts on airway responsiveness, inflammation, and remodeling in ovalbumin (OVA)-induced asthmatic mice. Mice with ovalbumin (OVA)-induced allergic asthma were treated with S. miltiorrhiza extracts, and airway resistance (RL) to methacholine, inflammatory cell infiltration, Th1/Th2 cytokine levels, and airway remodeling were assessed. TGF-β1-induced BEAS-2B and MRC-5 cells were used to evaluate the effects of five S. miltiorrhiza compounds on epithelial-mesenchymal transition and fibrosis. OVA-challenge resulted in remarkably increased RL, inflammatory cell infiltration, Th1/Th2 cytokine levels in BALF, goblet cell hyperplasia, collagen deposition, and airway wall thickening. Daily treatment with S. miltiorrhiza ethanolic (EE, 246 mg/kg) or water (WE, 156 mg/kg) extract significantly reduced OVA-induced airway inflammatory cell infiltration, Th1/Th2 cytokine amounts, and goblet cells hyperplasia. However, only WE remarkably decreased RL, collagen deposition, and airway wall thickening. Moreover, Chromatography showed that salvianic acid A and caffeic acid levels were much higher in WE than EE, while rosmarinic acid was slightly lower; salvianolic acid B and tanshinone IIA levels were much higher in EE than WE. Interestingly, caffeic acid and rosmarinic acid were more potent in reducing E-cadherin and vimentin levels in TGF-β1-induced BEAS-2B cells, and α-SMA and COL1A1 amounts in TGF-β1-induced MRC-5 cells. Both S. miltiorrhiza WE and EE alleviate airway inflammation in mice with OVA-sensitized allergic asthma. S. miltiorrhiza WE is more potent in reducing responsiveness and airway remodeling.

Luo J ，Zhang L ，Zhang X ，Long Y ，Zou F ，Yan C ，Zou W ... -  《-》

Experimental protocol for development of adjuvant-free murine chronic model of allergic asthma.

Mouse models of allergic asthma play a crucial role in exploring of asthma pathogenesis and testing of novel anti-inflammatory drugs. Widely used acute asthma models usually developed with adjuvant (aluminum hydroxide (alum)) do not reproduce one of the main asthma feature - airway remodeling while chronic asthma model mimic the pathophysiology of human disease. Moreover, the use of alum causes distress in experimental animals and impedes the test of adjuvant-containing drugs. In this study, we aimed to develop a chronic adjuvant-free asthma model with pronounced asthmatic phenotype. Female BALB/c mice were divided into 3 groups. The first group was sensitized with intraperitoneal injections of ovalbumin (OVA) emulsified in aluminum hydroxide on days 0, 14, 28 followed by two stages of intranasally challenge with OVA on days 41-43 and 62-64. The second group was subcutaneously sensitized with the same dose of OVA without adjuvant and challenged on the same days. The third group (negative control) included mice which did not received any kind of treatment (i.e. sensitization and challenge). Serum levels of OVA-specific IgE, IgG2a and IgG1 antibodies were detected by ELISA. Airway hyper-responsiveness was measured by non-invasive plethysmography on days 44 and 65. Bronchoalveolar lavage fluids (BALF) sampled in all groups on days 45 and 66 were analyzed by light microscopy. The left lung was removed for histological analysis. The IL-4 and IFNγ mRNA expression in BALF cells was evaluated by RT-PCR. The OVA-specific IgE antibody response was two-fold increased in mice from adjuvant-free group compared to the adjuvant group that reflects reorientation of immune response towards Th2 phenotype. At the same time, the level of OVA-specific IgG1 and IgG2a antibodies was increased in the adjuvant group. Airway hyperresponsiveness to methacholine in mice of both experimental groups was two-fold higher than in control. Analysis of cell composition in BAL has shown a significant increase in eosinophil count in both experimental groups that indicate the development of allergic inflammation. Lung histology revealed airway remodeling in both experimental groups including goblet cell hyperplasia/metaplasia, thickening of airway walls, collagen deposition in the wall of distal airways. Additionally, the tendency to develop hypertrophy of bronchial smooth muscle layer was observed. Study of gene expression in BAL cells revealed the increase of IL-4 level in both adjuvant and adjuvant-free groups while IFNγ expression in both experimental groups was similar to control group. We have developed a chronic adjuvant-free mouse asthma model which possesses all necessary features of the disease including airway remodeling and is more suitable for pre-clinical evaluation of novel therapeutic approaches including adjuvant-containing drugs.

Shilovskiy IP ，Sundukova MS ，Babakhin АА ，Gaisina AR ，Maerle AV ，Sergeev IV ，Nikolskiy AA ，Barvinckaya ED ，Kovchina VI ，Kudlay DA ，Nikonova AA ，Khaitov MR ... -  《-》