Neurogranin expression regulates mitochondrial function and redox balance in endothelial cells.

Autor: Jorgensen AN; Department of Pharmacology, Toxicology, and Neuroscience, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Rashdan NA; Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Rao KNS; Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Delgadillo LF; Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Kolluru GK; Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Krzywanski DM; Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Pattillo CB; Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Kevil CG; Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA., Nam HW; Department of Pharmacology, Toxicology, and Neuroscience, Louisiana State University Health Sciences Center, Shreveport, LA, 71103, USA. Electronic address: hyung.nam@lsuhs.edu.
Jazyk: angličtina
Zdroj: Redox biology [Redox Biol] 2024 Apr; Vol. 70, pp. 103085. Date of Electronic Publication: 2024 Feb 11.
DOI: 10.1016/j.redox.2024.103085
Abstrakt: Endothelial dysfunction and endothelial activation are common early events in vascular diseases and can arise from mitochondrial dysfunction. Neurogranin (Ng) is a 17kD protein well known to regulate intracellular Ca 2+ -calmodulin (CaM) complex signaling, and its dysfunction is significantly implicated in brain aging and neurodegenerative diseases. We found that Ng is also expressed in human aortic endothelial cells (HAECs), and depleting Ng promotes Ca 2+ -CaM complex-dependent endothelial activation and redox imbalances. Endothelial-specific Ng knockout (Cre-CDH5-Ng f/f ) mice demonstrate a significant delay in the flow-mediated dilation (FMD) response. Therefore, it is critical to characterize how endothelial Ng expression regulates reactive oxygen species (ROS) generation and affects cardiovascular disease. Label-free quantification proteomics identified that mitochondrial dysfunction and the oxidative phosphorylation pathway are significantly changed in the aorta of Cre-CDH5-Ng f/f mice. We found that a significant amount of Ng is expressed in the mitochondrial fraction of HAECs using western blotting and colocalized with the mitochondrial marker, COX IV, using immunofluorescence staining. Seahorse assay demonstrated that a lack of Ng decreases mitochondrial respiration. Treatment with MitoEbselen significantly restores the oxygen consumption rate in Ng knockdown cells. With the RoGFP-Orp1 approach, we identified that Ng knockdown increases mitochondrial-specific hydrogen peroxide (H 2 O 2 ) production, and MitoEbselen treatment significantly reduced mitochondrial ROS (mtROS) levels in Ng knockdown cells. These results suggest that Ng plays a significant role in mtROS production. We discovered that MitoEbselen treatment also rescues decreased eNOS expression and nitric oxide (NO) levels in Ng knockdown cells, which implicates the critical role of Ng in mtROS-NO balance in the endothelial cells.
Competing Interests: Declaration of competing interest Declarations of interest: none.
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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