In order to explore the variation of driver’s physiological and psychological indicators in driving different levels of highway, a series of real car dynamic experimental is carried out to test the drivers on four categories highway(freeway, first-class highway, second-class highway and third-class highway) utilizing NeXus-10, which is a 10 channel physiological monitoring and a battery powered biofeedback device, intended to monitor and feed back physiological parameters on the human body for relaxation training, muscle reeducation and prescription use. Through analyzing and discussing the physiological characteristics indicators datum (EEG, HR, BVP, RSP, etc.) which are collected by experiment, the dynamic variation law of six types of drivers’ physiological indicators is got on the different levels of highway. The conclusions provide experimental evidence for the highway alignment design, traffic facilities design and set, transportation environmental governance, etc.. It also provides some theoretical reference for the highway traffic management and control.