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Glucosamine Inhibits TNF‐α‐Induced Expression of Inflammatory Mediators In Rat Aortic Smooth Muscle Cells through Inhibition of NFκB Activation
Author(s) -
XING DONGQI,
Feng Wenguang,
Chen YiuFai,
Chatham John C,
Oparil Suzanne
Publication year - 2008
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.22.1_supplement.744.4
Subject(s) - western blot , chemokine , tumor necrosis factor alpha , phosphorylation , glucosamine , chemistry , iκbα , inflammation , nf κb , blot , microbiology and biotechnology , adhesion , cell adhesion molecule , vascular smooth muscle , biochemistry , biology , signal transduction , endocrinology , immunology , receptor , smooth muscle , gene , organic chemistry
Objective: This study tested whether glucosamine (GlcN), an amino suger that increases O‐GlcNAc modification of proteins by increasing flux through the hexosamine biosynthesis pathway, can protect vascular smooth muscle cells (SMCs) against inflammatory stress. Methods and Results: Quiescent cultured rat aortic SMCs (RASMCs) were pretreated with GlcN (5 mM) or vehicle for 1 hr and then stimulated with tumor necrosis factor (TNF)‐α (10 ng/ml) for another 15 min or 6 hrs. Total and phosphorylated IκBα were measured by western blot analysis. NFκB DNA binding activity were determined by ELISA. RNA was extracted for real‐time RT‐PCR analysis of chemokine and adhesion molecule expression. Nucleocytoplasmic protein subjected to western blot analysis of global O‐GlcNAcylated protein using the selective CTD110.6 antibody. GlcN treatment enhanced overall O‐GlcNAcylated protein levels and inhibited chemokine and adhesion molecule expression, IκBα phosphorylation, prevented the degradation of IκBα and inhibited NFκB activation in TNF‐α‐stimulated RASMCs. Conclusions: O‐GlcNAc modification may protect the vasculature from inflammatory stresses. This work was supported, in part, by American Heart Association Greater Southeast Affiliate grant 0425455B, 0765398B.