K(ATP) channel in vascular smooth muscle cell (VSMC) is closely linked to the etiology of hypertension. The aim of this study was to investigate effect of the high-fat diet-induced obesity on K(ATP) channel and blood pressure. Obesity was induced by a 24-week high-fat diet feeding in rats. Function and expression of K(ATP) channel in mesenteric arteries were examined using myography system, patch clamp and Western blotting. We show that high-fat diet increased blood pressure, decreased K(ATP) channel-mediated relaxation responses and currents, and down-regulated K(ATP) expression in VSMC. In conclusion, diet-induced obesity impairs K(ATP) channels in VSMC, which may underscore obesity-triggered increase in blood pressure.