Characterization of endothelial NADPH oxidase 5 effects on atherothrombotic stroke
Cardiovascular diseases (CVDs) are the leading cause of death globally, with ischemic stroke being a major concern. An ischemic stroke occurs approximately every three seconds. One key cause of ischemic stroke is atherosclerotic plaque, which can block blood vessels or rupture, leading to a thrombot...
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| Tipo de recurso: | tesis doctoral |
| Fecha de publicación: | 2024 |
| País: | España |
| Institución: | Consejo Superior de Investigaciones Científicas (CSIC) |
| Repositorio: | Dadun. Depósito Académico Digital de la Universidad de Navarra |
| Idioma: | inglés |
| OAI Identifier: | oai:dadun.unav.edu:10171/123823 |
| Acceso en línea: | https://hdl.handle.net/10171/123823 |
| Access Level: | acceso embargado |
| Palabra clave: | Cultivo celular Fisiología cardiovascular Arterioesclerosis y Arteroesclerosis Trombosis |
| Sumario: | Cardiovascular diseases (CVDs) are the leading cause of death globally, with ischemic stroke being a major concern. An ischemic stroke occurs approximately every three seconds. One key cause of ischemic stroke is atherosclerotic plaque, which can block blood vessels or rupture, leading to a thrombotic response and forming a thrombus that further occludes vessels. Oxidative stress, caused by an imbalance between the production and elimination of reactive oxygen species (ROS), is a primary molecular mechanism in atherothrombotic stroke. NADPH oxidases (NOXs) are enzymes that produce ROS, playing both physiological and pathological roles. Among the NOX family, NOX5 is unique, being regulated by intracellular calcium levels and not requiring complementary subunits to produce ROS. Although less studied due to its absence in rodent genomes, NOX5 is implicated in processes contributing to CVDs, including endothelial dysfunction and immune cell adhesion. NOX5 expression is elevated in atherosclerotic plaques and is increased by smoking. However, the detailed role of NOX5 in atherothrombotic stroke remains underexplored. We hypothesized that NOX5 contributes to atherothrombotic stroke through endothelial dysfunction, affecting processes such as extracellular matrix (ECM) modifications, thrombosis, blood-brain barrier (BBB) impairment, immune infiltration, and stroke onset. Our study aimed to uncover new molecular mechanisms and pathological processes driven by endothelial NOX5 in atherothrombotic stroke. To investigate, we used various models to examine the effects of NOX5 overexpression in endothelial cells. In vitro, we employed human aortic endothelial cells (teloHAEC), human brain microvascular endothelial cells (hCMEC/D3), and human umbilical vein endothelial cells (HUVEC), using adenovirus and expression plasmids for NOX5 overexpression. In vivo, we used a knock-in mouse model expressing human NOX5 in endothelial cells upon tamoxifen induction, subjecting these mice to FeCl3-induced carotid thrombosis and ischemic stroke. Additionally, in silico analyses were conducted to study proteomic changes in hCMEC/D3 cells infected with NOX5 adenovirus and thrombi from the carotid thrombosis model. Our results demonstrated that NOX5 overexpression in endothelial cells influences multiple stages of atherothrombotic stroke. Firstly, chronic NOX5 overexpression induces MMP-10 secretion, linked to ECM changes, atherosclerotic plaque instability, and stroke severity. Secondly, NOX5 promotes thrombosis through a sex-dependent mechanism: in females, via PGE2 secretion leading to inflammation, and in males, by activating neutrophils. Thirdly, NOX5 overexpression in BBB endothelial cells causes dysfunction, increasing apoptosis, cell migration, mitochondrial dysfunction, and reducing cell proliferation. Finally, NOX5 enhances immune cell infiltration into the brain by disrupting tight junctions and increasing immune cell adhesion, exacerbated by ischemic stroke. In conclusion, endothelial NOX5 plays a significant role in the pathophysiology of atherothrombotic ischemic stroke, acting as a pathological contributor or as a response to vascular damage. This identifies NOX5 as a potential therapeutic target for preventing atherothrombotic ischemic stroke. |
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