Logo image
Back
Critical role of NADPH oxidase-derived reactive oxygen species in generating Ca2+ oscillations in human aortic endothelial cells stimulated by histamine
Journal article   Open access   Peer reviewed

Critical role of NADPH oxidase-derived reactive oxygen species in generating Ca2+ oscillations in human aortic endothelial cells stimulated by histamine

Qinghua Hu, Zu-Xi Yu, Victor J Ferrans, Kazuyo Takeda, Kaikobad Irani and Roy C Ziegelstein
The Journal of biological chemistry, Vol.277(36), pp.32546-32551
09/06/2002
DOI: 10.1074/jbc.M201550200
PMID: 12093794
url
https://doi.org/10.1074/jbc.M201550200View
Published (Version of record) Open Access

Abstract

There is increasing evidence that intracellular reactive oxygen species (ROS) play a role in cell signaling and that the NADPH oxidase is a major source of ROS in endothelial cells. At low concentrations, agonist stimulation of membrane receptors generates intracellular ROS and repetitive oscillations of intracellular Ca(2+) concentration ([Ca(2+)](i)) in human endothelial cells. The present study was performed to examine whether ROS are important in the generation or maintenance of [Ca(2+)](i) oscillations in human aortic endothelial cells (HAEC) stimulated by histamine. Histamine (1 microm) increased the fluorescence of 2',7'-dihydrodichlorofluorescin diacetate in HAEC, an indicator of ROS production. This was partially inhibited by the NADPH oxidase inhibitor diphenyleneiodonium (DPI, 10 microm), by the farnesyltransferase inhibitor H-Ampamb-Phe-Met-OH (2 microm), and in HAEC transiently expressing Rac1(N17), a dominant negative allele of the protein Rac1, which is essential for NADPH oxidase activity. In indo 1-loaded HAEC, 1 microm histamine triggered [Ca(2+)](i) oscillations that were blocked by DPI or H-Ampamb-Phe-Met-OH. Histamine-stimulated [Ca(2+)](i) oscillations were not observed in HAEC lacking functional Rac1 protein but were observed when transfected cells were simultaneously exposed to a low concentration of hydrogen peroxide (10 microm), which by itself did not alter either [Ca(2+)](i) or levels of inositol 1,4,5-trisphosphate (Ins-1,4,5-P(3)). Thus, histamine generates ROS in HAEC at least partially via NADPH oxidase activation. NADPH oxidase-derived ROS are critical to the generation of [Ca(2+)](i) oscillations in HAEC during histamine stimulation, perhaps by increasing the sensitivity of the endoplasmic reticulum to Ins-1,4,5-P(3).
 Endothelium, Vascular - cytology Reactive Oxygen Species Histamine - pharmacology Signal Transduction Calcium - metabolism Humans Cells, Cultured Enzyme Inhibitors - pharmacology Hydrogen Peroxide - pharmacology NADPH Oxidases - metabolism Endoplasmic Reticulum - metabolism Onium Compounds - pharmacology Genes, Dominant Time Factors Endothelium, Vascular - metabolism Aorta - cytology Kinetics Inositol 1,4,5-Trisphosphate - metabolism

Details

Metrics

29 Record Views
72 Times Cited - Web of Science
Logo image