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Nitric oxide metabolites in induced sputum: a marker of airway inflammation in asthmatic subjects

Authors


Jang Assistant Professor, Department of Allergy and Pulmonology, Seonam University College of Medicine, Seogu Marukdong 120–1, Kwangju, Korea, 502–157.

Abstract

Background and objective

The role of nitric oxide (NO) needs to be further clarified in allergic inflammation. This study was designed to investigate the relationships between NO metabolites and eosinophil count, eosinophil cationic protein (ECP), interleukin (IL)-5 in induced sputum from asthmatics.

Methods

Hypertonic saline-induced sputum was obtained in 25 asthmatic subjects, among which 13 patients were examined before and after anti-asthmatic medications including steroid preparations. Ten normal subjects were enrolled as controls. Fresh expectorated sputum separated from saliva was treated with equal volume of dithiothreitol 0.1%, cytospined for cell count, and the supernatant was collected for biochemical assay. NO metabolites were assayed by using modified Griess reaction. ECP was measured by fluoroimmunoassay, and detected IL-5 by a sandwich ELISA.

Results

Asthmatic subjects, compared with controls, had significantly higher concentration of NO metabolites (1035.4 ± 125.3 vs 557.2 ± 101.5 μmol/L, P < 0.01), higher percentage of eosinophils (25.6 ± 4.6 vs 1.7 ± 0.2%, P < 0.01), and higher levels of ECP (1117.8 ± 213.9 vs 154.6 ± 47.4 μg/L, P < 0.01) in the induced sputum. IL-5 was detected more frequently in asthmatic subjects than in control subjects (11/25 [44%] vs 1/10 [10%], P < 0.05). According to asthma severity, moderate to severe asthmatic subjects (n = 18) had higher level of NO metabolites (1143.8 ± 156.3 vs 575.5 ± 89.5 μmol/L, P < 0.01), higher levels of ECP and IL-5 (P < 0.01, respectively) in the induced sputum than in those of mild asthmatic subjects (n = 7). NO metabolites, the percentage of eosinophils, the levels of ECP, and IL-5 were reduced following treatment with anti-asthmatic drugs (P < 0.01, respectively). There were significant positive correlations between NO metabolites and percentage of eosinophils or ECP (r = 0.34, P < 0.05; r = 0.28, P < 0.05). Negative correlations were noted between FEV1, FEV1/FVC and proportion of eosinophils, ECP, or IL-5 levels.

Conclusion

These findings confirmed that the level of NO metabolites was increased in the tracheobronchial secretion of asthmatic subjects and was paralleled with severity of asthma. Measurement of NO metabolites in induced sputum may be used for monitoring the degree of airway inflammation in asthmatics.

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