Purpose <p>This study aimed to examine the relieving effect of static stretching on cognitive fatigue derived from cognitive load with visual display terminal (VDT), and its psychophysiological and neurological mechanisms.</p> Methods <p>Thirty healthy university students performed 3-minute static stretching (ST condition) or seated rest (RE condition) on separate days. After cognitive load, the Color-ward Stroop test (CWST) was performed before and after stretching or rest (Pre/Post); psychological responses were evaluated after stretching or rest. Heart rate variability, pupil diameter, and prefrontal cortex activity were measured during the CWST before and after stretching or rest.</p> Results <p>The ST condition increased vitality (P &lt; 0.001), subjective arousal (P &lt; 0.001), and muscle tension relief (P &lt; 0.001) and improved executive function (EF) (P = 0.02) compared with the RE condition. Furthermore, EF improvement was correlated with stretching frequency (P = 0.03). Although physiological parameters did not differ between the conditions, pupil diameter and prefrontal cortex activity were&#xa0;correlated with subjective arousal (P = 0.02, P = 0.03).</p> Conclusion <p>Static stretching has a relieving effect on cognitive fatigue caused by cognitive load with VDT, and this effect could be modulated by stretching habits.</p>