NADPH Oxidase 5 Induces Changes in the Unfolded Protein Response in Human Aortic Endothelial Cells and in Endothelial-Specific Knock-in Mice.

Adriana Cortés,Juan José Martínez-Irujo,Carlos De Miguel,Silvia Cenoz,Íñigo Izal,Guillermo Zalba,Álvaro Pejenaute,Javier Marqués,Eduardo Ansorena

doi:10.3390/antiox10020194

Adriana Cortés, Juan José Martínez-Irujo + Show 7 more

Open Access

https://doi.org/10.3390/antiox10020194

Copy DOI

Abstract

Oxidative stress constitutes a key molecular mechanism in the development of cardiovascular diseases. A potential relationship between reactive oxygen species (ROS) driven by the NADPH oxidase family (NOX) and the unfolded protein response (UPR) has been postulated. Nevertheless, there is a lack of information about the crosstalk between NOX5 homologue and the UPR in a cardiovascular context. The main aim was to analyze NOX5-mediated ROS effects in the UPR and its importance in cardiovascular diseases. To this effect, we used an adenoviral NOX5-β overexpression model in human aortic endothelial cells (HAEC) and a conditional endothelial NOX5 knock-in mouse. Using expression arrays, we investigated NOX5-induced genomic changes in HAEC. Compared with the control HAEC, 298 genes were differentially expressed. Gene ontology analysis revealed the activation of numerous cellular routes, the most relevant being the UPR pathway. Using real-time PCR and Western Blot experiments, we confirmed that NOX5 overexpression induced changes in the expression of the UPR components, which were associated with increased apoptosis. Moreover, in endothelial-specific NOX5 knock-in mice, we found changes in the expression of the UPR components genes. In these mice, myocardial infarction was performed by permanent coronary artery ligation; however, NOX5 expression was not associated with differences in the UPR components mRNA levels. In these animals, we found significant associations between the UPR components gene expression and echocardiographic parameters. Our data support the idea that NOX5-derived ROS may modulate the UPR pathway in endothelial cells, which might play a relevant role in cardiac physiology.

Highlights

Cardiovascular diseases (CVDs) are the leading cause of death in the developed world, accounting for a 44% of non-communicable disease deaths [1]
We studied the unfolded protein response (UPR) components protein expression to better characterize NOX5-β implications in this pathway
human aortic endothelial cells (HAEC) infection with NOX5-β adenovirus produced different expression patterns in the protein levels of UPR-related genes, depending on the time of infection compared with green fluorescent protein (GFP)-infected cells (MOI 100) (Figure 4)

Summary

Introduction

Cardiovascular diseases (CVDs) are the leading cause of death in the developed world, accounting for a 44% of non-communicable disease (non-transmissible diseases) deaths [1]. Increased reactive oxygen species (ROS) production contributes to many aspects of pathological progression [3]. High production of ROS and attenuation of endogenous antioxidants promote an oxidative stress environment that precedes CVDs [4]. ROS can oxidize proteins, DNA and other cellular components causing cell damage [5]. All cellular types of the cardiovascular system produce ROS under physiological conditions, including endothelial cells, cardiomyocytes, smooth muscle cells and fibroblasts [6]. Their production increases under pathological conditions [7]

Methods

Results

Discussion

Conclusion