gw-501516 and ruboxistaurin

Activity of the vascular large conductance Ca(2+)-activated K(+) (BK) channel is tightly regulated by its accessory β(1) subunit (BK-β(1)). Downregulation of BK-β(1) expression in diabetic vessels is associated with upregulation of the forkhead box O subfamily transcription factor-3a (FOXO-3a)-dependent F-box-only protein (FBXO) expression. However, the upstream signaling regulating this process is unclear. Overproduction of reactive oxygen species (ROS) is a common finding in diabetic vasculopathy. We hypothesized that ROS signaling cascade facilitates the FOXO-3a/FBXO-mediated BK-β(1) degradation and leads to diabetic BK channel dysfunction. Using cellular biology, patch clamp, and videomicroscopy techniques, we found that reduced BK-β(1) expression in streptozotocin (STZ)-induced diabetic mouse arteries and in human coronary smooth muscle cells (SMCs) cultured with high glucose was attributable to an increase in protein kinase C (PKC)-β and NADPH oxidase expressions and accompanied by attenuation of Akt phosphorylation and augmentation of atrogin-1 expression. Treatment with ruboxistaurin (a PKCβ inhibitor) or with GW501516 (a peroxisome proliferator-activated receptor δ activator) reduced atrogin-1 expression and restored BK channel-mediated coronary vasodilation in diabetic mice. Our results suggested that oxidative stress inhibited Akt signaling and facilitated the FOXO-3a/FBXO-dependent BK-β(1) degradation in diabetic vessels. Suppression of the FOXO-3a/FBXO pathway prevented vascular BK-β(1) degradation and protected coronary function in diabetes.

Topics: Animals; Arteries; Cells, Cultured; Diabetes Mellitus, Experimental; Enzyme Inhibitors; F-Box Proteins; Forkhead Box Protein O3; Forkhead Transcription Factors; Glucose; Indoles; Large-Conductance Calcium-Activated Potassium Channel beta Subunits; Maleimides; Mice; Microscopy, Video; Muscle Proteins; Muscle, Smooth, Vascular; NADPH Oxidases; Patch-Clamp Techniques; Protein Kinase C; Protein Kinase C beta; Proteolysis; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; SKP Cullin F-Box Protein Ligases; Thiazoles; Vasodilation