Methotrexate (MTX) is a chemotherapeutic medicine frequently used to treat various forms of cancer. The objective of our research was to examine whether allicin (ALC) and lycopene (LP) could alleviate the harmful effects of MTX-induced testicular toxicity in rats. A total of 49 male white albino rats were divided into 7 groups (7 rats in each group). The rats of group 1 was kept as a control group and received normal saline orally; while rats of group 2 and group 3 administered ALC (20mg/kg/BW) LP (10mg/kg/BW) orally. On day 15, the rats of group 4 received a single dose of MTX (20mg/kg, IP). In group 5, rats received ALC and MTX; in group 6, rats received LP and MTX, and in group 7, rats received ALC, LP, and MTX. Results showed that MTX induced significant increase in values of testicular malondialdehyde (MDA) levels with a substantial reduction in glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and lowered the serum testosterone, follicle-stimulating hormone (FSH), and lutenizing hormone (LH) levels. MTX induced a marked decline in seminiferous tubule diameter and epithelium height. MTX showed a widening of the interstitial space of seminiferous tubules, atrophy, vacuolar degeneration of the germinal epithelium and Leydig's interstitial cells had scanty cytoplasm and vesicular nuclei. Also, testicular proliferating cell nuclear antigen (PCNA) expression was recorded. In MTX-intoxicated rats, ALC and LP increased antioxidant biomarkers, decreased MDA, and attenuated changes in serum hormonal parameters. It is concluded that ALC and LP have significant protective effects on MTX- induced testicular toxicity due to their antioxidant properties.