Platelet-derived extracellular vesicles express NADPH oxidase-1 (Nox-1), generate superoxide and modulate platelet function

Autor: Joanne L. Mitchell, Jonathan M. Gibbins, Giordano Pula, Renato Simões Gaspar, Plinio Ferreira
Rok vydání: 2021
Předmět:
Blood Platelets
Platelets
0301 basic medicine
Short Communication
CRP
collagen-related peptide
ERK
extracellular signal-regulated kinases
GAPDH
glyceraldehyde 3-phosphate dehydrogenase
NADPH Oxidase
Biochemistry
Collagen receptor
PRP
platelet-rich plasma
Extracellular Vesicles
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
NTA
nanoparticle tracking analysis
PMA
phorbol-12-myristate-13-acetate
Superoxides
Physiology (medical)
PKC
protein kinase C
EV
extracellular vesicles
TRAP-6
thrombin receptor activator peptide 6
Platelet
Platelet activation
Redox biology
chemistry.chemical_classification
Reactive oxygen species
NADPH oxidase
biology
Chemistry
Superoxide
NADPH Oxidases
Fibrinogen binding
NADPH
nicotinamide adenine dinucleotide phosphate
Platelet Activation
Cell biology
WP
washed platelets
030104 developmental biology
GPVI
Glycoprotein VI
PDEVs
platelet-derived extracellular vesicles
NADPH Oxidase 1
biology.protein
GPVI
Reactive Oxygen Species
030217 neurology & neurosurgery
Zdroj: Free Radical Biology & Medicine
ISSN: 0891-5849
DOI: 10.1016/j.freeradbiomed.2021.01.051
Popis: Background Platelets release platelet-derived extracellular vesicles (PDEVs) upon activation – in a process that is regulated by generation of reactive oxygen species (ROS). Platelet NADPH oxidase-1 (Nox-1) contributes to ROS generation and thrombus formation downstream of the collagen receptor GPVI. Objectives We aimed to investigate whether PDEVs contain Nox-1 and whether this is relevant for PDEV-induced platelet activation. Methods PDEVs were isolated through serial centrifugation after platelet activation with thrombin receptor agonist TRAP-6 (activated PDEVs) or in the absence of agonist (resting PDEVs). The physical properties of PDEVs were analyzed through nanoparticle tracking analysis. Nox-1 levels, fibrinogen binding and P-selectin exposure were measured using flow cytometry, and protein levels quantified by immunoblot analysis. ROS were quantified using DCF fluorescence and electron paramagnetic resonance. Results Nox-1 was found to be increased on the platelet outer membrane upon activation and was present in PDEVs. PDEVs induced platelet activation, while co-addition of GPVI agonist collagen-related peptide (CRP) did not potentiate this response. PDEVs were shown to be able to generate superoxide in a process at least partially mediated by Nox-1, while Nox-1 inhibition with ML171 (also known as 2-APT) did not influence PDEV production. Finally, inhibition of Nox-1 abrogated PDEV-mediated platelet activation. Conclusions PDEVs are able to generate superoxide, bind to and activate platelets in a process mediated by Nox-1. These data provide novel mechanisms by which Nox-1 potentiates platelet responses, thus proposing Nox-1 inhibition as a feasible strategy to treat and prevent thrombotic diseases.
Graphical abstract Image 1
Highlights • Activated platelets have increased NADPH oxidase-1 (Nox-1) exposure on the outer membrane. • Platelet-derived extracellular vesicles (PDEVs) express Nox-1 and generate superoxide in a process mediated by Nox-1. • PDEVs bind and activate platelets in a Nox-1-dependent manner.
Databáze: OpenAIRE
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