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Am. J. Respir. Cell Mol. Biol., Volume 24, Number 3, March, 2001 295-303

Expression of Heme Oxygenase in Human Airway Epithelial Cells

Louise E. Donnelly and Peter J. Barnes

Department of Thoracic Medicine, Imperial College School of Science, Technology and Medicine, National Heart and Lung Institute, London, United Kingdom

Elevated levels of carbon monoxide (CO) are found in the exhaled breath of patients with inflammatory diseases such as asthma and cystic fibrosis. Endogenous CO is derived from heme oxygenase (HO) (EC 1.14.99.3), which catabolizes heme-producing CO and biliverdin. There are three isoforms of HO: HO-1 is inducible by inflammatory cytokines and oxidants, including nitric oxide (NO), whereas HO-2 and HO-3 are expressed constitutively. Primary airway epithelial cells were treated with either 50 ng/ml interleukin-1beta , tumor necrosis factor-alpha , and interferon-gamma (cytomix), or the NO donor NOC-18 for up to 24 h. Cytomix-induced HO-1 expression peaked at 4 h, returning to baseline by 24 h, whereas HO-2 expression remained unchanged. This increase in HO-1 expression could not be explained by an increase in NO production as inducible NO synthase expression increased between 12 and 24 h. However, the NO donor NOC-18 (500 µM) increased HO-1 expression twofold and HO activity 25-fold, whereas cytomix treatment increased HO activity eightfold. NO induction of HO-1 was not mediated via guanylyl cyclase and was not attenuated by 1 µM dexamethasone, although dexamethasone increased HO-2 protein. Therefore, airway epithelial cells express HO-2 and can express HO-1; thus, the epithelium may be a source of increased CO in airway diseases.

Abbreviations: cyclic guanosine monophosphate, cGMP; carbon monoxide, CO; deoxynucleotide triphosphate, dNTP; enhanced chemiluminescence, ECL; fetal calf serum, FCS; granulocyte macrophage colony-stimulating factor, GM-CSF; Hanks' balanced salt solution, HBSS; heme oxygenase, HO; interleukin, IL; inducible nitric oxide synthase, iNOS; messenger RNA, mRNA; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT; nitric oxide, NO; nitric oxide synthase, NOS; reverse transcription/polymerase chain reaction, RT-PCR; sodium dodecyl sulfate, SDS; tumor necrosis factor, TNF.




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