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

Autor(es):Mostrar menos -

da Silva, Josiane Fernandes ^[1] ; Alves, V, Juliano ; Silva-Neto, Julio A. ^[2] ; Costa, Rafael M. ^[2] ; Neves, Karla B. ^[3] ; Alves-Lopes, Rheure ^[3] ; Camargo, Livia L. ^[3] ; Rios, Francisco J. ^[3] ; Montezano, Augusto C. ^[3] ; Touyz, Rhian M. ^[3] ; Tostes, Rita C. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo FMRP USP USP, Ribeirao Preto Med Sch, Dept Pharmacol, Sao Paulo - Brazil

^[2] Alves, Juliano, V, Univ Sao Paulo FMRP USP USP, Ribeirao Preto Med Sch, Dept Pharmacol, Sao Paulo - Brazil

^[3] Univ Glasgow, Inst Cardiovasc & Med Sci, BHF Glasgow Cardiovasc Ctr, Glasgow, Lanark - Scotland

• Tipo de documento: Artigo Científico
• Fonte: Clinical Science; v. 135, n. 15, p. 1845-1858, AUG 2021.
• Citações Web of Science: 0

Resumo

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. (AU)

• Processo FAPESP: 13/08216-2 - CPDI - Centro de Pesquisa em Doenças Inflamatórias
• Beneficiário: Fernando de Queiroz Cunha
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 17/26072-9 - Papel do ambiente pró-oxidativo e da oxidação de proteínas tirosina fosfatase em processos associados a aterosclerose
• Beneficiário: Josiane Fernandes da Silva
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado