Protein Kinase C-[beta] Contributes to Impaired Endothelial Insulin Signaling in Humans With Diabetes Mellitus.
Tabit, Corey E. MD, MBA; Shenouda, Sherene M. PhD; Holbrook, Monica MS; Fetterman, Jessica L. PhD; Kiani, Soroosh MD; Frame, Alissa A. BS; Kluge, Matthew A. MA; Held, Aaron MA; Dohadwala, Mustali M. MD; Gokce, Noyan MD; Farb, Melissa G. PhD; Rosenzweig, James MD; Ruderman, Neil MD, DPhil; Vita, Joseph A. MD; Hamburg, Naomi M. MD, MS
127(1):86-95, January 1, 2013.
(Format: HTML, PDF)
Background-: Abnormal endothelial function promotes atherosclerotic vascular disease in diabetes. Experimental studies indicate that disruption of endothelial insulin signaling, through the activity of protein kinase C-[beta] (PKC[beta]) and nuclear factor [kappa]B, reduces nitric oxide availability. We sought to establish whether similar mechanisms operate in the endothelium in human diabetes mellitus.
Methods and Results-: We measured protein expression and insulin response in freshly isolated endothelial cells from patients with type 2 diabetes mellitus (n=40) and nondiabetic controls (n=36). Unexpectedly, we observed 1.7-fold higher basal endothelial nitric oxide synthase (eNOS) phosphorylation at serine 1177 in patients with diabetes mellitus (P=0.007) without a difference in total eNOS expression. Insulin stimulation increased eNOS phosphorylation in nondiabetic subjects but not in diabetic patients (P=0.003), consistent with endothelial insulin resistance. Nitrotyrosine levels were higher in diabetic patients, indicating endothelial oxidative stress. PKC[beta] expression was higher in diabetic patients and was associated with lower flow-mediated dilation (r=-0.541, P=0.02). Inhibition of PKC[beta] with LY379196 reduced basal eNOS phosphorylation and improved insulin-mediated eNOS activation in patients with diabetes mellitus. Endothelial nuclear factor [kappa]B activation was higher in diabetes mellitus and was reduced with PKC[beta] inhibition.
Conclusions-: We provide evidence for the presence of altered eNOS activation, reduced insulin action, and inflammatory activation in the endothelium of patients with diabetes mellitus. Our findings implicate PKC[beta] activity in endothelial insulin resistance.
(C) 2013 American Heart Association, Inc.