Reactive oxygen species induced by diamide inhibit insulin-induced ATP-sensitive potassium channel activation in cultured vascular smooth muscle cells

Both insulin resistance and reactive oxygen species (ROS) have been reported to play essential pathophysiological roles in cardiovascular diseases. However, the mechanistic link between ROS and insulin resistance in the vasculature remains unclear. Recently we have shown that insulin causes K _ATP channel activation mediated by PI3K in cultured vascular smooth muscle cells (VSMCs). K_ATP channel in VSMCs is critical in the regulation of vascular tonus. Here we examined the effects of ROS induced by a thol-oxidizing agent, diamide, on the insulin signalling pathway and K _ATP channel activities in cultured VSMCs (A10 cells). Diamide (100 μM) increased intercellular ROS and extracellular signal-regulated kinases (ERK) activitiy. Treatment with 100 μM diamide suppressed significantly insulin-induced IRS and Akt phosphorylation. In addition to IRS and Akt, diamide inhibited insulin receptor auto-phosphorylation. Patch-clamp study showed that diamide suppressed insulin-induced but did not pinacidil-induced K_ATP channel activities in A10 cells. From these data, we conclude that ROS inhibit critical insulin signal transduction components including IRS and Akt, and these effects cause down-regulation of insulin's action in the vasculature including K_ATP channel activation. This study may contribute to our understanding of mechanisms of insulin resistance-associated cardiovascular disease.