The endothelial dysfunction is considered to be an initial step in atherosclerosis. Additionally, it was reported that the smooth muscle, which constructs the media of the artery, changes its characteristics owing to atherosclerosis. Therefore, it is essential to develop a method of assessing the regional endothelial function and mechanical properties of the arterial wall. To evaluate the endothelial function, a conventional technique of measuring the transient change in the diameter of the brachial artery caused by flow-mediated dilation (FMD) after the release of avascularization is used. However, this method can not evaluate the mechanical properties of the wall. We previously developed a method for the simultaneous measurements of waveforms of radial strain and blood pressure in the radial artery. In this study, the viscoelasticity of the arterial wall was estimated from the measured stress-strain relationship using the least-squares method and the transient changes in the mechanical properties of the arterial wall ware revealed. From in vivo experimental results, the stress-strain relationship showed a hysteresis loop and viscoelasticity was estimated by the proposed method. The slope of the loop decreased owing to FMD, which resulted in the decrease in estimated elastic modulus. The increase in the area of the loop occurred after recirculation, which corresponds to the increase in the ratio of the loss modulus (depends on viscosity) to the elastic modulus when the Voigt model is assumed. In this study, the variance in estimates was evaluated by in vivo measurement for 10 min. The temporal decrease in static elasticity after recirculation due to FMD was much larger than the evaluated variance. These results show a potential of the proposed method for the thorough analysis of the transient change in viscoelasticity due to FMD.