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Modulatory effects of N-acetyl-L-cysteine on human eosinophil apoptosis

Depts of ¹ Pharmacology, and ³ Medicine, University of Valencia, ² Research Foundation, and, ⁴ Service of Pneumology, Consortium University General Hospital, and ⁵ Clinical Pharmacology Unit, University Clinic Hospital, Valencia, Spain.

CORRESPONDENCE: E. J. Morcillo, Dept of Pharmacology, Faculty of Medicine, Av. Blasco Ibáñez 15, E-46010 València, Spain. Fax: 34 63864622. E-mail: Esteban.Morcillo{at}uv.es

Keywords: Apoptosis, cytokines, human eosinophils, N-acetyl-L-cysteine

Received: June 5, 2006
Accepted May 4, 2007

Eosinophils are oxidant-sensitive cells considered relevant in allergic inflammation. The present study aimed to examine the effects of the antioxidant N-acetyl-L-cysteine (NAC) on constitutive and cytokine-delayed apoptosis in human isolated eosinophils.

Human eosinophils were purified from the blood of healthy donors by a magnetic separation system. Apoptosis and cellular glutathione were assessed by cytofluorometric analysis and nuclear factor (NF)-B binding activity assessed by electrophoresis mobility shift assay.

The rate of spontaneous apoptosis of human eosinophils after 24 h culture, as assessed by annexin-V-positive staining, was mean±sem 48.2±1.4%, n = 5. Granulocyte-macrophage colony-stimulating factor (GM-CSF; 10 ng·mL^–1) decreased apoptosis to 19.4±1.8%, n = 5. NAC (5 mM) inhibited spontaneous apoptosis (33.6±2.7%, n = 5) but augmented apoptosis in the presence of GM-CSF (30.9±1.5%, n = 5). NAC (5 mM) also increased the rate of apoptosis in the presence of tumour necrosis factor (TNF)- (10 ng·mL^–1) and interleukin-5 (5 ng·mL^–1). NAC (5 mM) increased eosinophil glutathione content. The increase in eosinophil NF-B binding activity induced by GM-CSF and TNF- was suppressed by NAC.

In conclusion, N-acetylcysteine modulates eosinophil apoptosis by inhibiting constitutive apoptosis but reversing the survival effect produced by inflammatory cytokines in human eosinophils.

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