Abstract: Fatigue effect is one of the factors leading to the damage of chemical film on optical components in high power laser system. However, few attentions are given on this issue. In this work, two irradiation types, single fluences with multi-shots and multi-gradient fluence with multi-shots, are utilized to investigate the characteristics and effect of laser fatigue damage on antireflection film at 1064 nm. The results indicate that fatigue damage is easy to occur on film under multi-shot laser irradiation. While the damage threshold and the resistance of fatigue effect can be effectively improved at the multi-gradient fluence with multi-shots. The results can provide references on methods and technique to improve the resistance ability and resist the fatigue damage of film.