Expression and activity of nitric oxide synthases in human airway epithelium
| Autor: | Kerrie A Basclain, Philip J. Thompson, D.N. Watkins, M.J. Garlepp, D.J. Peroni |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Pulmonary and Respiratory Medicine
Pathology medicine.medical_specialty Clinical Biochemistry Bronchi In situ hybridization Nitric Oxide Polymerase Chain Reaction Epithelium Gene Expression Regulation Enzymologic Nitric oxide chemistry.chemical_compound medicine Humans RNA Messenger Cloning Molecular Enzyme Inhibitors Molecular Biology In Situ Hybridization Nitrites Cell Line Transformed biology Epithelial Cells Cell Biology Blotting Northern Molecular biology Nitric oxide synthase Blotting Southern Carcinoma Bronchogenic NG-Nitroarginine Methyl Ester medicine.anatomical_structure Thoracotomy chemistry Cell culture biology.protein Cytokines Respiratory epithelium Immunohistochemistry Tumor necrosis factor alpha Nitric Oxide Synthase Histamine |
| Zdroj: | American Journal of Respiratory Cell and Molecular Biology. 16:629-639 |
| ISSN: | 1535-4989 1044-1549 |
| DOI: | 10.1165/ajrcmb.16.6.9191464 |
| Popis: | Nitric oxide (NO) synthesized by airway epithelium may be important in the regulation of airway inflammation and reactivity. As such, the expression and localization of nitric oxide synthase (NOS) isoforms was assessed in human airway tissue obtained following thoracotomy, and in cultured human airway epithelial (BEAS-2B) cells. NOS expression was determined by reverse transcription-polymerase chain reaction (RT-PCR)/Southern blot analysis, and localized by in situ hybridization and immunohistochemistry. No synthesis by cell cultures was detected by nitrite assay. Endothelial and neuronal constitutive NOS mRNAs were not detected in airways or cell cultures. Inducible NOS (iNOS) mRNA was detected in 5 of 6 airway specimens, and in situ hybridization demonstrated iNOS mRNA expression in columnar epithelial cells. This was confirmed by immunohistochemistry using an iNOS specific antibody. BEAS-2B cell cultures were stimulated with (I) combinations of tumor necrosis factor alpha (TNF alpha) (50-2,000 U/ml)/interferon gamma (IFN gamma) (20-500 U/ml)/lipopolysaccharide (LPS) (10 micrograms/ml) or (2) histamine (10(-3) M-10(-5) M). Cell cultures treated with TNF alpha/IFN gamma/LPS in combination expressed iNOS mRNA, and this was associated with increases in supernatant nitrite concentrations over 24 h; however, this response diminished with successive passage of cells. Histamine treatment did not result in iNOS mRNA expression or detectable NO synthesis. We conclude that iNOS in human airway tissue is localized to the airway epithelium. Cytokine/ LPS stimulation, but not histamine, results in iNOS mRNA expression and NO synthesis in BEAS-2B cells. BEAS-2B cells may not be a suitable model for the study of NO biology in airway epithelium. |
| Databáze: | OpenAIRE |
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