The cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP triggered chloride channel in the apical membrane of epithelial cells, is malfunctioning in patients with cystic fibrosis (CF). Research attempts are centered on chloride channel function in order to discover a remedy for the disease. Genistein increased chloride transport in normal and delF508-CFTR cultured airway epithelial cells without cAMP stimulation. Prior pretreatment with phenylbutyrate didn’t impact the rate of the genistein-stimulated chloride efflux in these cells. S-nitrosoglutathione is an endogenous bronchodilator, found in reduced quantities in the lungs of CF patients…
Contents: Functional Aspects of Epithelia in Cystic Fibrosis and Asthma
Introduction
Cystic Fibrosis
Cystic Fibrosis Transmembrane Conductance Regulator
CF pathophysiology
The airways
Clinical background
Airway epithelium
Ion transport in airway epithelium
Sweat glands
CF diagnosis, survival and prognosis
Pharmacological treatment of CF
Genistein
S-nitrosoglutathione (GSNO)
Other agents
Asthma
Airway inflammation and remodeling in asthma
ICAM
Treatment of asthma
Corticosteroids
Leukotriene modifiers
2. Aims
3. Methods
3.1 Cells
3.1.1 Cultured cell lines (Papers I, II, III, IV, V)
3.1.2 Nasal epithelial cells (Paper II)
3.2 Transmission electron microscopy
3.3 Protein expression
3.3.1 Immunoblot analysis (Paper II)
3.3.2 Immunocytochemistry (Paper IV)
3.3.3 Flow cytometry (Paper V)
3.4 CFTR function
3.4.1 X-ray microanalysis (Paper I,II)
3.4.2 Intracellular chloride measurements (Paper I, I, III, IV)
3.5 Intracellular calcium measurements (Paper II)
3.6 Transepithelial resistance (TEER) (Paper III)
3.7 Detection of apoptosis and necrosis (Paper V)
3.8 ICAM expression (Paper V)
3.9 Statistical analysis
Results
4.1 Paper I
4.2 Paper II
4.3 Paper III
4.4 Paper IV
4.5 Paper V…
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