Acetaminophen (APAP), a widely used medication known for its pain-relieving and fever-reducing effects, can cause kidney failure if taken in excess. To investigate the potential protective effects of allicin (ALC) and/or omega-3 fatty acids (O3FA) against acetaminophen-induced kidney damage, a study was conducted using 49 rats divided into seven groups. The control group was given saline, while the other groups received ALC, O3FA, APAP, ALC + APAP, O3FA + APAP, or ALC + O3FA + APAP. After administering APAP, the rats showed decreased levels of total protein and albumin in their blood, along with increased levels of creatinine and urea. The concentration of reduced glutathione (GSH), as well as the activity of superoxide dismutase (SOD) and catalase (CAT), decreased, while the level of malondialdehyde (MDA) in the renal tissues increased. The activation of caspase-3 and HSP70 also suggested an impact on kidney histopathology. Overall, the study found that ALC and/or O3FA may have a protective impact against acetaminophen-induced kidney damage through their anti-inflammatory, anti-apoptotic, and antioxidant defense systems.