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Abstract 462: Ascl3 Protects Vascular Smooth Muscle Cells Against Oxidative Stress

Swarajit K Biswas, Liliana Hernandez, Yan Xiao, Mushtaq Ahmad, Minerva T Garcia-Barrio
Arteriosclerosis, Thrombosis, and Vascular Biology. 2013;33:A462
Swarajit K Biswas
Cardiovascular Rsch Institute, Morehouse Sch of Medicine, Atlanta, GA
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Liliana Hernandez
Cardiovascular Rsch Institute, Morehouse Sch of Medicine, Atlanta, GA
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Yan Xiao
Cardiovascular Rsch Institute, Morehouse Sch of Medicine, Atlanta, GA
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Mushtaq Ahmad
Cardiovascular Rsch Institute, Morehouse Sch of Medicine, Atlanta, GA
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Minerva T Garcia-Barrio
Cardiovascular Rsch Institute, Morehouse Sch of Medicine, Atlanta, GA
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Abstract

Atherosclerosis initiation and progression depends on multiple risk factors and is sustained by self-perpetuating inflammatory cellular and molecular events that ultimately translate into increased oxidative stress inducing endothelial dysfunction, macrophage activation and altering vascular smooth muscle cell (VSMC) homeostasis. Our initial work identified the under-characterized bHLH achaete-scute like 3, ASCL3, as a pro-proliferative anti-apoptotic factor in VSMC and prompted the hypothesis that ASCL3 plays a protective role against oxidative stress in VSMC. We determined that ASCL3 expression is up-regulated in the media and lesions of the atherosclerotic aorta in ApoE-mice fed a high fat diet. In vitro, ASCL3 expression is consistent with an adaptive role in oxidative stress. In primary RASMC, time- and dose-response to H2O2 (0.5-2mM, 4-24h) result in marked ASCL3 mRNA increase (~10-fold) at low concentrations or shorter times and is reduced as oxidative stress increases. siRNA knockdown of ASCL3 increases susceptibility of RASMC to 0.25mM H2O2 (low dose) as determined by caspase-3 activation. Stable over-expression of ASCL3 in A7r5 cells results in enhanced protection at higher doses of H2O2, and is associated with higher AKT and ERK1/2 phosporylation. This protective effect involves the mitochondrial pathway since ASCL3 up-regulates Bcl-2/Bax ratio in basal and induced conditions (0.4mM, 4h), inhibits mitochondrial depolarization, prevents mitochondrial fragmentation and preserves mitochondrial DNA. ROS levels are significantly reduced in ASCL3 over-expressing cells (0.2mM, 2h). ASCL3 over-expression appears to increase mitochondrial density and results in altered mitochondrial and cristae morphology as determined by TEM. Surprisingly, subcellular localization studies indicate that ASCL3 localizes primarily to the mitochondria. In summary, low oxidative stress in VSMC promotes survival, with enhanced proliferation, and as it increases, induces adaptation associated with an antioxidant gene expression profile. Eventually high oxidative stress leads to cell death. Altogether, our data indicate that ASCL3 plays a protective role in the survival and adaptation of vascular smooth muscle cells to oxidative stress.

  • Oxidative stress
  • ASCL3 bHLH
  • mitochondria
  • © 2013 by American Heart Association, Inc.
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April 2018, Volume 38, Issue 4
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    Abstract 462: Ascl3 Protects Vascular Smooth Muscle Cells Against Oxidative Stress
    Swarajit K Biswas, Liliana Hernandez, Yan Xiao, Mushtaq Ahmad and Minerva T Garcia-Barrio
    Arteriosclerosis, Thrombosis, and Vascular Biology. 2013;33:A462, originally published October 20, 2015

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    Abstract 462: Ascl3 Protects Vascular Smooth Muscle Cells Against Oxidative Stress
    Swarajit K Biswas, Liliana Hernandez, Yan Xiao, Mushtaq Ahmad and Minerva T Garcia-Barrio
    Arteriosclerosis, Thrombosis, and Vascular Biology. 2013;33:A462, originally published October 20, 2015
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