Nicotinamide adenine dinucleotide phosphate reduced oxidase 5 (Nox5) regulation by angiotensin II and endothelin-1 is mediated via calcium/calmodulin-dependent, rac-1-independent pathways in human endothelial cells

Montezano, A. C.; Burger, Dylan; Paravicini, T. M.; Chignalia, A. Z.; Yusuf, Hiba; Almasri, Mahmoud; He, Ying; Callera, G. E.; He, Gang; Krause, Karl-Heinz; Lambeth, David; Quinn, M. T.; Touyz, R. M.

doi:10.1161/CIRCRESAHA.109.216036

Advanced search

Browse by...

Deposit

Highlights

More informations

Title		Nicotinamide adenine dinucleotide phosphate reduced oxidase 5 (Nox5) regulation by angiotensin II and endothelin-1 is mediated via calcium/calmodulin-dependent, rac-1-independent pathways in human endothelial cells
Authors		Montezano, A. C. Burger, Dylan Paravicini, T. M. Chignalia, A. Z. Yusuf, Hiba Almasri, Mahmoud He, Ying Callera, G. E. He, Gang Krause, Karl-Heinz Lambeth, David Quinn, M. T. Touyz, R. M. show all authors [1 - 13]
Published in		Circulation research. 2010, vol. 106, no. 8, p. 1363-1373
Abstract		RATIONALE: Although Nox5 (Nox2 homolog) has been identified in the vasculature, its regulation and functional significance remain unclear. OBJECTIVES: We sought to test whether vasoactive agents regulate Nox5 through Ca(2+)/calmodulin-dependent processes and whether Ca(2+)-sensitive Nox5, associated with Rac-1, generates superoxide (O(2)(-)) and activates growth and inflammatory responses via mitogen-activated protein kinases in human endothelial cells (ECs). METHODS AND RESULTS: Cultured ECs, exposed to angiotensin II (Ang II) and endothelin (ET)-1 in the absence and presence of diltiazem (Ca(2+) channel blocker), calmidazolium (calmodulin inhibitor), and EHT1864 (Rac-1 inhibitor), were studied. Nox5 was downregulated with small interfering RNA. Ang II and ET-1 increased Nox5 expression (mRNA and protein). Effects were inhibited by actinomycin D and cycloheximide and blunted by diltiazem, calmidazolium and low extracellular Ca(2+) ([Ca(2+)](e)). Ang II and ET-1 activated NADPH oxidase, an effect blocked by low [Ca(2+)](e), but not by EHT1864. Nox5 knockdown abrogated agonist-stimulated O(2)(-) production and inhibited phosphorylation of extracellular signal-regulated kinase (ERK)1/2, but not p38 MAPK (mitogen-activated protein kinase) or SAPK/JNK (stress-activated protein kinase/c-Jun N-terminal kinase). Nox5 small interfering RNA blunted Ang II-induced, but not ET-1-induced, upregulation of proliferating-cell nuclear antigen and vascular cell adhesion molecule-1, important in growth and inflammation. CONCLUSIONS: Human ECs possess functionally active Nox5, regulated by Ang II and ET-1 through Ca(2+)/calmodulin-dependent, Rac-1-independent mechanisms. Nox5 activation by Ang II and ET-1 induces ROS generation and ERK1/2 phosphorylation. Nox5 is involved in ERK1/2-regulated growth and inflammatory signaling by Ang II but not by ET-1. We elucidate novel mechanisms whereby vasoactive peptides regulate Nox5 in human ECs and demonstrate differential Nox5-mediated functional responses by Ang II and ET-1. Such phenomena link Ca(2+)/calmodulin to Nox5 signaling, potentially important in the regulation of endothelial function by Ang II and ET-1.
Keywords		Angiotensin II/ metabolism — Calcium/ metabolism — Calcium Channel Blockers/pharmacology — Calmodulin/antagonists & inhibitors/ metabolism — Cells, Cultured — Diltiazem/pharmacology — Endothelial Cells/drug effects/ enzymology — Endothelin-1/ metabolism — Enzyme Activation — Enzyme Inhibitors/pharmacology — Gene Expression Regulation, Enzymologic — Humans — Imidazoles/pharmacology — Inflammation/enzymology — JNK Mitogen-Activated Protein Kinases/metabolism — Membrane Proteins/genetics/ metabolism — Mitogen-Activated Protein Kinase 1/metabolism — Mitogen-Activated Protein Kinase 3/metabolism — NADPH Oxidase/genetics/ metabolism — Phosphorylation — Proliferating Cell Nuclear Antigen/metabolism — Pyrones/pharmacology — Quinolines/pharmacology — RNA Interference — RNA, Messenger/metabolism — Signal Transduction/drug effects — Superoxides/metabolism — Time Factors — Vascular Cell Adhesion Molecule-1/metabolism — p38 Mitogen-Activated Protein Kinases/metabolism — rac1 GTP-Binding Protein/antagonists & inhibitors/ metabolism
Identifiers		DOI: 10.1161/CIRCRESAHA.109.216036 PMID: 20339118
Full text		Article - Limited access to UNIGE Other version: http://circres.ahajournals.org/cgi/reprint/106/8/1363.pdf
Citation (ISO format)		MONTEZANO, A. C. et al. Nicotinamide adenine dinucleotide phosphate reduced oxidase 5 (Nox5) regulation by angiotensin II and endothelin-1 is mediated via calcium/calmodulin-dependent, rac-1-independent pathways in human endothelial cells. In: Circulation research, 2010, vol. 106, n° 8, p. 1363-1373. doi: 10.1161/CIRCRESAHA.109.216036 https://archive-ouverte.unige.ch/unige:11479