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Sodium salicylate rescues inflammation‐associated decrease in phosphorylated‐eNOSser1177 in human aortic endothelial cells through an AMPK dependent mechanism (LB706)
Author(s) -
Siefers Kyle,
WegmanPoints Lauren,
Witmer Jordan,
Pierce Gary
Publication year - 2014
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.28.1_supplement.lb706
Subject(s) - enos , ampk , medicine , phosphorylation , endocrinology , chemistry , amp activated protein kinase , endothelial dysfunction , nitric oxide , protein kinase a , nitric oxide synthase , biochemistry
Obesity is associated with impaired nitric oxide (NO)‐mediated vascular endothelial function (VEF). Pro‐inflammatory cytokines, such as TNF, may contribute to reduced VEF in part through a direct reduction of endothelial NO synthase (eNOS) expression and/or phosphorylation. Sodium salicylate (NaSal), a non‐acetylated aspirin that inhibits nuclear factor B and activates AMP kinase (AMPK), improves VEF in obese humans but the molecular mechanisms remain unclear. We tested the hypothesis that NaSal increases eNOS expression/phosphorylation in TNF‐stimulated endothelial cells through an AMPK‐dependent mechanism. Human aortic endothelial cells (HAECs) incubated in vitro with TNFα (10 ng/ml, 2 hrs) demonstrated decreased (vs. control) expression (Western blotting) of eNOS serine 1177 phosphorylation (p‐eNOS ser1177 , n=6; P<0.01 ), eNOS (n=6, P=.078 ) and phosphorylated AMPK (p‐AMPK Thr172 , n=2, P<0.05 ). Pretreatment of HAECs with NaSal (5 mM, 30 min) prohibited the TNF‐stimulated decrease in eNOS (n=4; P=0.77 ), p ‐eNOS ser117 (n=4; P=0.1 ) and p‐AMPK Thr172 (n=2; P=0.22 ). Co‐treatment with the AMPK inhibitor compound C (10 μM, 30 min) abolished the TNF‐induced rescue of p‐eNOS ser1177 (n=2; P<0.05 ) but not eNOS (n=2, P=0.77 ) by NaSal. These preliminary data suggest that NaSal contributes to increased phosphorylated eNOS ser1177 in TNF‐stimulated endothelial cells in part through inhibiting AMPK. Grant Funding Source : Supported by the University of Iowa Fraternal Order of Eagles Diabetes Research Center pilot grant.