By factorial experiment, the frequency of blink and rate of time of closed eyes of the measuring subjects are analyzed to judge the degree of visual fatigue when they are in different time learning with the eyes of the electrical signal under the lighting environment of a combination of different color temperature and different shades of light. The results showed that:the interaction effects of illumination, color temperature, illumination time and light intensity have significant influence on frequency of blink and the rate of change in closed eye time. Among them, the color temperature and illumination has the greatest interaction effects. With the increasing of illumination, visual fatigue decreases firstly and then rises. The optimal level of illumination is 1 000~2 000 lx, and the high illumination of over 3 000 lx should be avoided. Under the condition of 4 000 k color temperature, the blink frequency and the rate of closed eyes time are both less and the visual fatigue is relatively low. With the increase of the time of illumination, the rate of blink frequency and the time of closed eyes increase and the visual fatigue aggravates. The optimum illumination time is 0.5~1 h and should not exceed 1.5 h.