Free Fatty Acid Impairment of Nitric Oxide Production in Endothelial Cells Is Mediated by IKK

Objective—Free fatty acids (FFA) are commonly elevated in diabetes and obesity and have been shown to impair nitric oxide (NO) production by endothelial cells. However, the signaling pathways responsible for FFA impairment of NO production in endothelial cells have not been characterized. Insulin receptor substrate-1 (IRS-1) regulation is critical for activation of endothelial nitric oxide synthase (eNOS) in response to stimulation by insulin or fluid shear stress. Methods and Results—We demonstrate that insulin-mediated tyrosine phosphorylation of IRS-1 and serine phosphorylation of Akt, eNOS, and NO production are significantly inhibited by treatment of bovine aortic endothelial cells with 100 mol/L FFA composed of palmitic acid for 3 hours before stimulation with 100 nM insulin. This FFA preparation also increases, in a dose-dependent manner, IKK activity, which regulates activation of NFB, a transcriptional factor associated with inflammation. Similarly, elevation of other common FFA such as oleic and linoleic acid also induce IKK activation and inhibit insulin-mediated eNOS activation. Overexpression of a kinase inactive form of IKK blocks the ability of FFA to inhibit insulin-dependent NO production, whereas overexpression of wild-type IKK recapitulates the effect of FFA on insulin-dependent NO production. Conclusions—Elevated levels of common FFA found in human serum activate IKK in endothelial cells leading to reduced NO production, and thus may serve to link pathways involved in inflammation and endothelial dysfunction. (Arterioscler Thromb Vasc Biol. 2005;25:989-994.)