Central role of c-Src in NOX5- mediated redox signalling in vascular smooth muscle cells in human hypertension
Camargo, Livia L. and Montezano, Augusto C. and Hussain, Misbah and Wang, Yu and Zou, Zhiguo and Rios, Francisco J. and Neves, Karla B. and Alves-Lopes, Rheure and Awan, Fazli R. and Guzik, Tomasz J. and Jensen, Thomas and Hartley, Richard C. and Touyz, Rhian M. (2022) Central role of c-Src in NOX5- mediated redox signalling in vascular smooth muscle cells in human hypertension. Cardiovascular Research, 118 (5). pp. 1359-1373. ISSN 0008-6363 (https://doi.org/10.1093/cvr/cvab171)
Preview |
Text.
Filename: Camargo_etal_CR_2021_Central_role_of_c_Src_in_NOX5_mediated_redox_signalling_in_vascular_smooth_muscle_cells.pdf
Final Published Version License: Download (1MB)| Preview |
Abstract
Aims: NOX-derived reactive oxygen species (ROS) are mediators of signalling pathways implicated in vascular smooth muscle cell (VSMC) dysfunction in hypertension. Among the numerous redox-sensitive kinases important in VSMC regulation is c-Src. However, mechanisms linking NOX/ROS to c-Src are unclear, especially in the context of oxidative stress in hypertension. Here, we investigated the role of NOX-induced oxidative stress in VSMCs in human hypertension focusing on NOX5, and explored c-Src, as a putative intermediate connecting NOX5-ROS to downstream effector targets underlying VSMC dysfunction. Methods and results: VSMC from arteries from normotensive (NT) and hypertensive (HT) subjects were studied. NOX1,2,4,5 expression, ROS generation, oxidation/phosphorylation of signalling molecules, and actin polymerization and migration were assessed in the absence and presence of NOX5 (melittin) and Src (PP2) inhibitors. NOX5 and p22phox-dependent NOXs (NOX1-4) were down-regulated using NOX5 siRNA and p22phox-siRNA approaches. As proof of concept in intact vessels, vascular function was assessed by myography in transgenic mice expressing human NOX5 in a VSMC-specific manner. In HT VSMCs, NOX5 was up-regulated, with associated oxidative stress, hyperoxidation (c-Src, peroxiredoxin, DJ-1), and hyperphosphorylation (c-Src, PKC, ERK1/2, MLC20) of signalling molecules. NOX5 siRNA reduced ROS generation in NT and HT subjects. NOX5 siRNA, but not p22phox-siRNA, blunted c-Src phosphorylation in HT VSMCs. NOX5 siRNA reduced phosphorylation of MLC20 and FAK in NT and HT. In p22phox- silenced HT VSMCs, Ang II-induced phosphorylation of MLC20 was increased, effects blocked by melittin and PP2. NOX5 and c-Src inhibition attenuated actin polymerization and migration in HT VSMCs. In NOX5 transgenic mice, vascular hypercontractilty was decreased by melittin and PP2. Conclusion: We define NOX5/ROS/c-Src as a novel feedforward signalling network in human VSMCs. Amplification of this system in hypertension contributes to VSMC dysfunction. Dampening the NOX5/ROS/c-Src pathway may ameliorate hypertension-associated vascular injury.
ORCID iDs
Camargo, Livia L., Montezano, Augusto C., Hussain, Misbah, Wang, Yu, Zou, Zhiguo, Rios, Francisco J., Neves, Karla B. ORCID: https://orcid.org/0000-0001-5158-9263, Alves-Lopes, Rheure, Awan, Fazli R., Guzik, Tomasz J., Jensen, Thomas, Hartley, Richard C. and Touyz, Rhian M.;-
-
Item type: Article ID code: 82758 Dates: DateEvent30 April 2022Published28 July 2021Published Online26 July 2021AcceptedSubjects: Medicine Department: Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences Depositing user: Pure Administrator Date deposited: 13 Oct 2022 14:12 Last modified: 03 Oct 2024 03:42 URI: https://strathprints.strath.ac.uk/id/eprint/82758