Lysophosphatidylcholine induces oxidative stress in human endothelial cells via NOX5 activation - implications in atherosclerosis

Da Silva, J. F. et al. (2021) Lysophosphatidylcholine induces oxidative stress in human endothelial cells via NOX5 activation - implications in atherosclerosis. Clinical Science, 135(15), pp. 1845-1858. (doi: 10.1042/CS20210468) (PMID:34269800)

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Objective: The mechanisms involved in NOX5 activation in atherosclerotic processes are not completely understood. The present study tested the hypothesis that lysophosphatidylcholine (LPC), a proatherogenic component of oxLDL, induces endothelial calcium influx, which drives NOX5-dependent reactive oxygen species (ROS) production, oxidative stress, and endothelial cell dysfunction. Approach: Human aortic endothelial cells (HAEC) were stimulated with LPC (10−5 M, for different time points). Pharmacological inhibition of NOX5 (Melittin, 10−7 M) and NOX5 gene silencing (siRNA) was used to determine the role of NOX5-dependent ROS production in endothelial oxidative stress induced by LPC. ROS production was determined by lucigenin assay and electron paramagnetic spectroscopy (EPR), calcium transients by Fluo4 fluorimetry, and NOX5 activity and protein expression by pharmacological assays and immunoblotting, respectively. Results: LPC increased ROS generation in endothelial cells at short (15 min) and long (4 h) stimulation times. LPC-induced ROS was abolished by a selective NOX5 inhibitor and by NOX5 siRNA. NOX1/4 dual inhibition and selective NOX1 inhibition only decreased ROS generation at 4 h. LPC increased HAEC intracellular calcium, important for NOX5 activation, and this was blocked by nifedipine and thapsigargin. Bapta-AM, selective Ca2+ chelator, prevented LPC-induced ROS production. NOX5 knockdown decreased LPC-induced ICAM-1 mRNA expression and monocyte adhesion to endothelial cells. Conclusion: These results suggest that NOX5, by mechanisms linked to increased intracellular calcium, is key to early LPC-induced endothelial oxidative stress and pro-inflammatory processes. Since these are essential events in the formation and progression of atherosclerotic lesions, the present study highlights an important role for NOX5 in atherosclerosis.

Item Type:Articles
Additional Information:This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo [FAPESP-CRID 2013/08216-2 (RCT); 2017/26072-9 (JFS), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil (RCT). This study was supported by grants from the British Heart Foundation (BHF) (RE18/6/34217). RMT is supported through a BHF Chair award (CH/12/29762) and ACM is supported through a Walton Fellowship, University of Glasgow.
Glasgow Author(s) Enlighten ID:Rios, Dr Francisco and Da Silva, Dr Josiane and Montezano, Dr Augusto and Alves Moreira Lopes, Dr Rheure and Touyz, Professor Rhian and Neves, Dr Karla
Creator Roles:
Da Silva, J.Conceptualization, Data curation, Formal analysis, Investigation, Writing – original draft, Writing – review and editing
Neves, K.Writing – review and editing
Lopes, R.Writing – review and editing
Rios, F.Writing – review and editing
Montezano, A.Writing – review and editing
Touyz, R.Writing – review and editing
Authors: Da Silva, J. F., Alves, J. V., Neto, J., Costa, R. M., Neves, K. B., Lopes, R., Carmargo, L., Rios, F., Montezano, A., Touyz, R. M., and Tostes, R. C.
College/School:College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health
Journal Name:Clinical Science
Publisher:Portland Press
ISSN (Online):1470-8736
Published Online:04 August 2021
Copyright Holders:Copyright © 2021 The Authors
First Published:First published in Clinical Science 135(15): 1845-1858
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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