Cyclophosphamide (CP) is a bifunctional alkylating and chemotherapeutic agent used to treat cancers. However, its clinical utilization is restricted due to its toxicity to many tissues, most notably the liver and kidneys. Four groups (n=6) of rats were used. Group I was given normal saline orally for 14 days. Group II glutathione (GSH) (150 mg/kg) orally once a day for 14 days. Group III was administered 0.5 ml normal saline orally once daily and injected cyclophosphamide on 10th day (200 mg/kg, intraperitoneal). Group IV received GSH (150 mg/kg) orally plus intraperitoneal CP (200 mg/kg). CP induced a significant increase in hepatic serum biomarkers including aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) levels compared with the control group. Additionally, there was a significant increase in total bilirubin (BIT). Also, levels of total protein (T-protein) and albumin (ALB) were significantly decreased in the CP group. Moreover, cyclophosphamide significantly decreased catalase (CAT) and glutathione (GSH) levels in liver tissues, while increasing malondialdehyde (MDA) levels. Interleukin-1β (IL- 1β) was substantially upregulated by CP in liver tissues. Furthermore, CP induced severe histopathological and immunochemical alteration in liver tissues and administering GSH concurrently with CP led to a notable enhancement in estimated parameters compared to the CP-only group. Due to its ability to safeguard against CP-induced liver damage in rats, GSH is advised for use.