To accurately and comprehensively explore the entire process of physical fatigue development in rescue team members during weighted walking, a multidimensional fatigue assessment method based on eye movement characteristics, electromyographic signals, and subjective evaluation is proposed. Eight volunteers were recruited for the weight-bearing walking fatigue induction experiment. The glasses eye tracking was used to extract the eye movement data about saccade time ( ST ), average saccade speed ( SS ) and maximum saccade amplitude ( SA ). The correlation between the characteristics of eye movement and the degree of fatigue estimated by subjective evaluation was -0.857 ± 0.059, -0.938 ± 0.092, not correlated, respectively. The correlation with iEMG to judge fatigue degree was -0.782 ± 0.090, -0.942 ± 0.030, -0.928 ± 0.026, respectively. Multiple linear regression analysis was performed on subjective score, iEMG value and eye movement parameters. The regression equations were Y=0.270X1-0.16X2-0.513X3-0.124X4+80.917, R2=0.989. This study makes new explorations and attempts in the monitoring and evaluation methods of fatigue during weighted walking.