Methotrexate (MTX) is an anti-folate used to treat cancer and some inflammatory diseases. The efficacy of MTX is often limited by its severe toxicity. Propolis is one of the most significant bee products that has an important role in balancing antioxidant systems and has an anti-peroxidant effect on several tissues. The present study was conducted to investigate the ameliorative effect of propolis against the methotrexate induced oxidative damage on blood, liver and kidney in rats. A total of one hundred and twenty male Wistar albino rats of mean body weights 90 ± 5 g were divided into 5 groups. Rats were administered their respective doses of propolis and/or MTX for 3, 6 and 9 weeks intervals. Control groups (G₁ and G₂), propolis group (G₃), methotrexate group (G₄) and methotrexate and propolis co-administered group (G₅).The results showed that the MTX significant reduced hemoglobin concentration, hematocrit %, MCHC value, RBCs, WBCs and platelets count and significantly increased MCV and lymphocytes %. Moreover, it caused a significant increase in MDA level and significant decrease in GSH concentration in liver tissue as compared to normal control group (G₁) in a time dependent manner. MTX administration also caused significant increase in serum AST, ALT and ALP activities in a time dependent manner, but a significant increase in total bilirubin only in 9 weeks as compared to normal control (G₁). On the other hand, MTX impaired kidney function as reflected by a significant increase in serum urea and creatinine levels and decrease in serum uric acid level as compared to normal control group. Results suggest that administration of propolis extract (G₅) help reduce the deleterious effects due to MTX injection in a time dependent manner, whereas, administration for 9 weeks offers better alleviation over 6 or 3 weeks. Data showed that long term administration of MTX for 9 weeks produce maximum damage over 6 or 3 weeks. Propolis extract administration also recovered the structural and functional integrity of the hepatic cells.