Asthma and Airway Inflammation
Asthma is a chronic inflammatory disorder arising from not fully understood heterogenic gene-environment interactions. Characteristic features include variable airway obstruction and bronchial hyperresponsiveness. It is frequently known to coexist with atopic diseases, particularly allergic rhinitis.
Human Airway Smooth Muscle cells stimulated with Poly I:C
The effect of PDE4 inhibitor on IL6 and IL8 release from Poly I:C stimulated Human Airway Smooth muscle cells (HASMs) was assessed. HASMs were pretreated with the PDE4 inhibitor (1 nM) for 1 hour prior to stimulation with Poly I:C (10 µg/ml) for 24 hours. Cell culture supernatants were then collected and analysed for IL6 and IL8 release by ELISA (n=3±SEM; **p < 0.01).
Human Lung Fibroblasts stimulated with Poly I:C
Primary human lung fibroblasts were treated with TL3 antagonists in conjunction with Poly I:C stimulation for 48 hours. The levels of IL-6, IL-8 and MCP-1 released in the supernatants were determined using Luminex Multiplex assay. One-way ANOVA was performed to determine statistical significance (*p<0.05; **p<0.01; ***p <0.001).
Human Lung Fibroblasts stimulated with TNFα
Primary human lung fibroblasts were treated with reference compound – Hydrocortisone in conjunction with TNF-α-mediated stimulation for 48 hours. Expression levels of IL1b, IL6, IL8, IFNγ, MMP1 and HMGB1 were determined using an ELISA-based assay. One-way ANOVA was performed to determine statistical significance (*p<0.05; **p<0.01; ***p <0.001).
Human Lung Fibroblasts stimulated with TNFα
Primary human lung fibroblasts were pre-treated with the test and reference compound (hydrocortisone) followed by stimulation with TNFα for 48 hours. Expression levels of PAI-1 and a-SMA were determined using an ELISA-based assay. TNFα stimulation upregulated PAI-1 and a-SMA levels and this was inhibited by hydrocortisone. (***p <0.001; n=3±SEM).
Human airway smooth muscle cells were embedded within collagen and pre-treated with 2,3 BDM followed by incubation for 4 hours with Carbachol. Images were captured every 30 mins over a period of 3 and a half hours. Percentage change in gel contraction was then determined. For each treatment the percentage change in the gel contraction was calculated based on the values obtained at time 0. Each condition is representative of 3 biological replicates (n=3); mean+SEM. Data analysis was performed using the 2-WAY ANOVA analysis, *p<0.05; **p<0.01, ***p<0.0001; Carbachol vs. test ingredients groups.
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