Methotrexate (MTX), a synthetic antimetabolite with a wide range of therapeutic indications, although its liver toxicity induced mainly through oxidative stress limits its clinical use. Vitamin C (ascorbic acid) possesses potent antioxidant and anti-inflammatory properties, Therefore, has a possible hepatoprotective effect. This study seeks to address the hepatoprotective effects of vitamin C against MTX-induced liver injury in albino mice. MTX showed significant elevation in both serum Alanine aminotransferase (ALT) and liver tissue Malondialdehyde (MDA), indicating hepatic injury, while vitamin C pre-treatment will hold down this elevation significantly and dose-dependently, causing amelioration of the toxic effect of MTX; the histopathological findings also support this finding. Also, MTX causes consumption of liver tissue content of superoxide dismutase (SOD) and Glutathione (GSH), while vitamin C pre-treatment boosts the SOD hepatic tissue level while GSH is diminished even more. In conclusion, vitamin C has a dose-dependent amelioration effect on the toxic effect of MTX on the liver.