Neutrophil-derived microparticles induce myeloperoxidase-mediated damage of vascular endothelial cells

BMC Cell Biology

Washington Luis Conrado dos-Santos¹  Alberto Augusto Noronha-Dutra²  Lain Carlos Pontes-de-Carvalho¹  Marilda Souza Gonçalves¹  Valéria Matos Borges¹  Thayna Meirelles³  Viviane Costa Junqueira Rocha¹  Luciana de Aragão França¹  Thassila Nogueira Pitanga⁴ 

[1]Fundação Oswaldo Cruz-BA, Gonçalo Moniz Research Center, Salvador, BA, Brazil

[2]University College London, Gower Street, London, UK

[3]Heart Institute, Faculty of Medicine, USP, São Paulo, SP, Brazil

[4]Health Science Institute, Federal University of Bahia, Salvador, BA, Brazil

关键词: Oxidative stress;    Myeloperoxidase;    Endothelial cell injury;    Neutrophil microparticles;   

Others  :  852111
DOI  :  10.1186/1471-2121-15-21

 received in 2013-12-21, accepted in 2014-06-05,  发布年份 2014

【 摘 要 】

Background

Upon activation neutrophil releases microparticles - small plasma membrane vesicles that contain cell surface proteins and cytoplasmic matter, with biological activities. In this study we investigated the potential role of myeloperoxidase in the endothelial cell injury caused by neutrophil-derived microparticles.

Results

Microparticles were produced by activating human neutrophils with a calcium ionophore and characterized by flow cytometry and transmission and scanning electron microscopy. Myeloperoxidase activity was measured by luminol-dependent chemiluminescence. Neutrophil microparticles-induced injuries and morphological alterations in human umbilical vein endothelial cells (HUVECs) were evaluated by microscopy and flow cytometry. Neutrophil microparticles were characterized as structures bounded by lipid bilayers and were less than 1 μm in diameter. The microparticles also expressed CD66b, CD62L and myeloperoxidase, which are all commonly expressed on the surface of neutrophils, as well as exposition of phosphatidylserine. The activity of the myeloperoxidase present on the microparticles was confirmed by hypochlorous acid detection. This compound is only catalyzed by myeloperoxidase in the presence of hydrogen peroxide and chloride ion. The addition of sodium azide or taurine inhibited and reduced enzymatic activity, respectively. Exposure of HUVEC to neutrophil microparticles induced a loss of cell membrane integrity and morphological changes. The addition of sodium azide or myeloperoxidase-specific inhibitor-I consistently reduced the injury to the endothelial cells. Taurine addition reduced HUVEC morphological changes.

Conclusions

We have demonstrated the presence of active myeloperoxidase in neutrophil microparticles and that the microparticle-associated myeloperoxidase cause injury to endothelial cells. Hence, the microparticle-associated myeloperoxidase-hydrogen peroxide-chloride system may contribute to widespread endothelial cell damage in conditions of neutrophil activation as observed in vasculitis and sepsis.

【 授权许可】

   
2014 Pitanga et al.; licensee BioMed Central Ltd.

【 图 表 】

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