In this study, it was aimed to investigate the effect of Esculetin on liver and kidney damage induced by methotrexate (MTX) in rats. Our study consisted of 4 groups: Group 1 (Control group), Group 2 (MTX), Group 3 (Esculetin) and Group 4 (MTX+Esculetin). Groups 3 and 4 received a dose of 100 mg / kg esculetin by orally during 14 days. In addition, groups 2 and 4 received MTX at a dose of 20 mg / kg intraperitoneally on the 9th day of the experiment. The study was terminated 5 days after the MTX application. Liver and kidney tissue MDA levels increased, GSH level and CAT and GSH-Px enzyme activities significantly decreased in MTX group and MTX administration caused oxidative stress in these tissues. Esculetin administration reduced the MDA level in liver and kidney tissues and increased the CAT and GSH-Px enzyme activities. In addition, esculetin administration decreased serum AST, ALT. At the histopathological examination, MTX caused degenerative and inflammatory changes such as hydropic degeneration and inflammatory cell infiltration in both liver and kidney. It was determined that esculetin significantly reduced these lesions. In conclusion, esculetin can prevent or minimize MTX induced liver and kidney damage due to its antioxidant and anti-inflammatory activities.