Respiratory Assays
COPD
The GOLD initiative defines COPD as “a disease state characterized by airflow limitation that is not fully reversible. The airflow limitation is usually both progressive and associated with an abnormal inflammatory response of the lungs to noxious particles or gases.” The American Thoracic Society defines COPD in terms of chronic bronchitis and emphysema. Chronic bronchitis is characterized by the clinical symptoms of excessive cough and sputum production; emphysema refers to chronic dyspnea, resulting from enlarged air spaces and destruction of lung tissue. COPD is the fourth leading cause of death worldwide.
Goblet cell hyperplasia
Human airway epithelial cells were differentiated into goblet cells and then treated with pulsed 5% cigarette smoke extract (CSE) and 1 µM Acrolein (continuous) for 28 days. Immunofluorescence staining for MUC5AC was performed at Day 14 and Day 28 and imaged using the Molecular Devices Xpress Pico machine.
MUC5AC, MUC5B, IL8 and FOXJ1 were analysed using Quantigene in samples stimulated with IL-13, IL-4, or unstimulated, over a period of 21 days. MUC5AC, MUC5B and IL8 all showed an increase in expression when stimulated with IL-13 or IL-4, whereas FOXJ1 showed a decrease in expression relative to the unstimulated control. Data are mean+SEM, N=6, analysed with one-way ANOVA, *p<0.05, ***p<0.001.
MUC5AC and MUC5B expression were measured using Quantigene analysis. An increase in mucin expression was observed in CSE and Acrolein treated cells relative to the day 2 untreated control. Data expressed as mean±SEM, N=9, analysed with one-way ANOVA, ***p<0.001.
Immunohistochemical analysis of MUC5AC and MUC5B in unstimulated and IL-13 stimulated samples. A. Unstimulated, B. IL-13 stimulated, both stained for MUC5AC. C. Unstimulated, D. IL-13 stimulated, both stained for MUC5B. Brown = positive cells, blue negative cells.
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