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Investigating the Effect of Inhibiting 'Reactive Oxygen Species' on Intracellular Signaling Pathways in Endothelial Cells
Author(s) -
Ashworth Isa,
Obeidat Yasin,
Puentes Laura,
Torina Anali,
Shafique Ehtesham,
Abid Ruhul
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.1046.3
Subject(s) - reactive oxygen species , pi3k/akt/mtor pathway , protein kinase b , signal transduction , microbiology and biotechnology , mapk/erk pathway , phosphorylation , intracellular , oxidative stress , chemistry , tumor necrosis factor alpha , foxo1 , blot , biology , biochemistry , immunology , gene
Objective Elevated Reactive Oxygen Species (ROS) in Endothelial Cells (EC's) have been implicated in the pathogenesis of cardiovascular diseases, and much research has gone into elucidating the mechanisms through which oxidative stress leads to these diseases. Research in the last decade, however, has uncovered the central role lower levels of ROS play in physiological intracellular signaling pathways, such as those stimulated by Vascular Endothelial Growth Factor (VEGF) and Tumour Necrosis Factor‐α (TNF‐α) in EC's. Our objective in this study was to determine the effect of a lack of ROS in EC's on the intracellular signaling pathways stimulated by VEGF and TNF‐α. Methods The activity of these pathways can be measured by the expression of key downstream proteins that are common to these two pathways such as Akt, ERK‐1/2, IKKα/B, AMPK, eNOS, mTOR and FOXO1. To investigate these pathways in vitro , Bovine Endothelial Aortic Cells (BAEC's) were treated with ROS inhibitors (NAC, DPI) and then exposed to VEGF or TNF‐α to stimulate their respective pathways. Protein was then extracted and transferred to a membrane by Western Blotting then visualized by immunoflourescence. Results Western blots demonstrated that reduction of ROS levels by NAC inhibited VEGF‐induced Akt phosphorylation but not phosphorylation of ERK1/2 in BAEC. The findings of TNF‐induced signaling activation were not conclusive. Conclusion Whereas VEGF‐mediated activation of PI3K‐Akt activation requires ROS, ERK1/2 activation is independent of ROS in ECs. This research was funded by National Institutes of Health (NIH) and American Heart Association (AHA) grants.