Hydrogen peroxide (H2O2) as an endothelium‐derived vasodilator factor mediates flow‐induced dilation (FID) in human adipose arterioles from patients with coronary artery disease (CAD). The intracellular sources responsible for H2O2 generation during FID remain unclear. This study is designed to test whether NADPH oxidase (NOX) serves as a potential source of reactive oxygen species (ROS) formation contributing to FID. Visceral adipose arterioles (~150 μm ID) obtained from patients with or without CAD were cannulated and pressurized at 60 mmHg for videomicroscopic measurement of vessel diameters. After constriction with endothelin‐1, arterioles were subjected to stepwise increase in luminal flow before and after treatment with gp91‐ ds‐tat, a cell‐permeable peptide inhibitor of NOX, or a scrambled control peptide. Flow induced a graded dilation of adipose arterioles from CAD (maximal dilation: 61±3%, n=6), and this dilation was markedly reduced by gp91‐ds‐tat (50μM; 5±4%, n=5, P<0.05). FID was not affected by the scrambled peptide (67±2 vs. 61±3% of controls). In contrast to arterioles from CAD patients, gp91‐ds‐tat had no effect on FID in arterioles from patients without CAD (68±2% vs. 67±2% of controls, n=5–6). In conclusion, NOX plays an essential role in FID of human adipose arterioles.