Inhibitory effects of N-acetylcysteine on the functional responses of human eosinophils in vitro

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Background: Oxidative stress appears to be relevant in the pathogenesis of inflammation in allergic diseases like bronchial asthma. Eosinophils are oxidant-sensitive cells considered as key effectors in allergic inflammation.

Objective: The aim of this work was to study the effects of the clinically used antioxidant N-acetyl-L-cysteine (NAC) on the functional responses of human-isolated eosinophils.

Methods: Human eosinophils were purified from the blood of healthy donors by a magnetic bead separation system. The effects of NAC were investigated on the generation of reactive oxygen species (chemiluminescence and flow cytometry), Ca(2+) signal (fluorimetry), intracellular glutathione (GSH; flow cytometry), p47(phox)-p67(phox) translocation (Western blot) and eosinophil cationic protein (ECP) release (radioimmunoassay).

Results: NAC (0.1-1 mm) inhibited the extracellular generation of oxygen species induced by N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and eotaxin (in the presence of IL-5) with -logIC(50) values of 3.61+/-0.03 and 3.36+/-0.09, respectively. Also, the intracellular generation of hydrogen peroxide was virtually abolished by NAC (0.5-1 mm). NAC (1 mm) did not alter the fMLP-induced Ca(2+) signal but augmented the eosinophil content of reduced GSH and inhibited p47(phox)-p67(phox) translocation. NAC inhibited the release of ECP ( approximately 90% inhibition at 1 mm) from fMLP-activated eosinophils.

Conclusion: Inhibition by NAC of human eosinophil functions in vitro is potentially useful in the treatment of allergic inflammation.

Source: Clinical and Experimental Allergy, 2007. May;37(5):714-22. PMID: 17456219, by Martinez-Losa M, Cortijo J, Juan G, O’Connor JE, Sanz MJ, Santangelo F, Morcillo EJ. Department of Pharmacology, University of Valencia, Valencia, Spain. [E-mail:
Esteban.Morcillo@uv.es]

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