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Zinc activates neutrophils’ oxidative burst

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Abstract

Zinc has been shown to disturb the innate host defense response by interfering in the activation of neutrophils and subsequent oxidative burst, although the exact role of this metal, either as an activator or inhibitor, remains a matter of controversy among research groups. These apparent discrepancies may be due to experimental settings, through modification of zinc availability to neutrophils, or to inaccurate detections of reactive species. Thus, the main objective of the present study was to provide clarification on the role of zinc on the activation of human neutrophils and the subsequent oxidative burst. For that purpose, different detection methods and incubation media were used. The obtained results showed that phosphate buffers (PBS and HBSS) complex with zinc and interfere with the results obtained with this metal. By using Tris-G, it was clearly demonstrated that zinc, at low concentrations (5–12.5 μM), activates NADPH oxidase, mainly via protein kinase C, leading to the formation of superoxide radical (O2 •−). Higher concentrations of zinc results on a rapid dismutation of O2 •− to oxygen and hydrogen peroxide, which in turn is used by myeloperoxidase to generate hypochlorous acid (HOCl).

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Abbreviations

PMN:

Polymorphonuclear leukocytes

ROS:

Reactive oxygen species

RNS:

Reactive nitrogen species

PKC:

Protein kinase C

ABAH:

4-Aminobenzoyl hydrazide

PMA:

Phorbol myristate acetate

PBS:

Dulbecco’s phosphate buffer saline

HBSS:

Hank’s balanced salt solution

Tris-G:

Tris buffer saline

APF:

6-(4′-Amino)phenoxy-3H-xanthen-3-on-9-yl]benzoic acid

HRP:

Horseradish peroxidase

Amplex Red:

10-Acetyl-3,7-dihydroxyphenoxazine

DPI:

Diphenyleneiodonium chloride

Gö6983:

3-[1-[3-(Dimethylamino)propyl]-5-methoxy-1H-indol-3-yl]-4-(1H-indol-3-yl)-1H-pyrrole-2,5-dione

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Acknowledgments

Marisa Freitas acknowledges Fundação para a Ciência e Tecnologia (FCT) and Fundo Social Europeu (FSE) her PhD grant (SFRH/BD/28502/2006).

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Authors and Affiliations

  1. REQUIMTE, Departamento de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha 164, 4099-030, Porto, Portugal

    Marisa Freitas, José L. F. C. Lima & Eduarda Fernandes

  2. Serviço de Hematologia Clínica, Hospital Geral de Santo António, Porto, Portugal

    Graça Porto

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  1. Marisa Freitas
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  2. Graça Porto
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  3. José L. F. C. Lima
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  4. Eduarda Fernandes
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Correspondence to Eduarda Fernandes.

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Freitas, M., Porto, G., Lima, J.L.F.C. et al. Zinc activates neutrophils’ oxidative burst. Biometals 23, 31–41 (2010). https://doi.org/10.1007/s10534-009-9264-x

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