Abstract Introduction: The most common side effects associated with the use of methotrexate (MTX) in the commonly used doses is hepatic toxicity. As a herbal alternative Hordeumvulgare seeds (Commonly known as Barley) commonly used by traditional medical practitioners in the treatment of many diseases including liver diseases.Aim of the work: The aim of this study was to study the potential protective effect ofBarley's grains on MTX induced liver injury. Materials and methods: adult male rats were used. The animals were divided into three groups each of them consists of 10 rats: Group I: (control group): were injected with IP saline . Group II: were IP injected withMTX in a dose of 6mg/kg /day for 5 days. Group III: Barley grains were given at a dose of 200g /kg/day for 30 days plusintraperitoneal injections of MTX on the day 25 , at the dose 6mg/kg /day for 5 days.the rats were sacrificed by decapitation then liver pieces for light microscopic study were obtained . Results: MTX markedly affected the liver tissue as appeared by light microscopic, immunohistochemical examinations and morphometric studies. Pretreatment of Barley's grains could attenuate some of these changes. Conclusion: Pretreatment of Barley's grains had a possible protective effect againstMTX induced liver injury.