Activation of IKKbeta by glucose is necessary and sufficient to impair insulin signaling and nitric oxide production in endothelial cells.
Hyperglycemic impairment of nitric oxide (NO) production by endothelial cells is implicated in the effect of diabetes to increase cardiovascular disease risk, but the molecular basis for this effect is unknown. In skeletal muscle, diabetes induces activation of inhibitor kappaB kinase (IKKbeta), a key cellular mediator of the response to inflammatory stimuli, and this impairs insulin signal transduction via the insulin receptor substrate-phosphatidylinositol 3-OH kinase (IRS-1/PI3-kinase) pathway. Since activation of endothelial nitric oxide synthase (eNOS) is dependent on IRS-1/PI3-kinase signaling, we hypothesized that activation of IKKbeta may contribute to the effect of glucose to impair NO production. Here, we show that exposure of bovine aortic endothelial cells to high glucose (25 mM) for 24 h impaired insulin-mediated tyrosine phosphorylation of IRS-1, serine phosphorylation of Akt, activation of eNOS, and production of NO. High glucose treatment also activated IKKbeta, and pretreatment with aspirin, a pharmacological inhibitor of IKKbeta, prevented both glucose-induced IKKbeta activation and the effect of high glucose to impair insulin-mediated NO production. These adverse responses to glucose were also blocked by selective inhibition of IKKbeta signaling via overexpression of a kinase-inactive form of the enzyme. Conversely, overexpression of wild-type IKKbeta recapitulated the deleterious effect of high glucose on insulin-mediated activation of eNOS. These data demonstrate that activation of IKKbeta plays a critical and novel role to mediate the deleterious effects of high glucose on endothelial cell function.
Department of Medicine, Harborview Medical Center, University of Washington, Box 359748, Seattle, WA 98104, USA. fkim@u.washington.edu
Francis
F
Kim
Kelly A
KA
Tysseling
Julie
J
Rice
Byron
B
Gallis
Lutfiyah
L
Haji
Cecilia M
CM
Giachelli
Elaine W
EW
Raines
Marshall A
MA
Corson
Michael W
MW
Schwartz
D000818:Animals; D001241:Aspirin / Q000494:pharmacology; D002417:Cattle; D042783:Endothelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D051550:I-kappa B Kinase / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
D007328:Insulin; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D051550:I-kappa B Kinase; D005947:Glucose; D001241:Aspirin