Photobiomodulation (PBM) has emerged as a potentially effective therapeutic approach to modulate cellular functions. This study aimed to examine the impact of PBM on reactive oxygen species (ROS) and lipid peroxidation, and glutathione-related antioxidant defense systems in rabbit eye tissues. A polychromatic light source with an intensity of 2.6 J/cm ²/min was used for PBM treatment in New Zealand White rabbits for 12 minutes. The PBM group (n=8) received treatments every two days for a total of twelve sessions, while the control group (n=8) did not undergo any PBM light exposure during the same period. The application of PBM significantly elevated ROS-mediated glutathione levels, along with increased activities of glutathione peroxidase and reductase, particularly in corneal tissue ( p ≤ 0.05). In conclusion, these findings could enhance understanding of PBM’s therapeutic potential for eye health and lead to new strategies for preventing or treating ocular diseases caused by oxidative stress.