Background: Pumpkin contains several phyto-constituents belonging to the categories of alkaloids, flavonoids, and palmitic, oleic and linoleic acids. Various important medicinal properties including anti-diabetic, antioxidant, anti-carcinogenic, anti-inflammatory and others have been well documented. Azathioprine (AZA), known as Imuran is animmunosuppressive drug. It is widely used in many diseases. A major drawback is the occurrence of side-effects, especially hepatorenal dysfunction. Objective: Assessing the association between the daily oral dose of pumpkin supplementation as a strategy for amelioration of the side-effects of AZA on liver functions, renal functions and some inflammatory markers and tumour necrosis factor-α. Material and Methods: Twenty four adult male albino rats, weighing 150–180 g, were used. They were divided into four equal groups: Group I (control), group II treated with (4 ml/kg. b. wt.) of pumpkin seeds oil per day for four weeks by oral gavage, group III were injected intraperitoneally with AZA (10mg/kg b. wt.) 3 times with an interval of 48 h in between, and group IV was treated by oral gavages with 4 ml/kg. b. wt of pumpkin seeds oil for ten consecutive days followed by AZA treatment i.p. at dose 10 mg/kg b. wt. 3 times with an interval of 48 h in between, after which pumpkin seeds oil administration alone was continued for another 2 weeks. Serum ALT, AST, ALP, total bilirubin, urea, creatinine, CRP and TNF-α were detected. Results: Pumpkin seeds oil supplementation with AZA treatment significantly decreased the levels of serum liver transaminases (ALT and AST), ALP and total bilirubin in addition to serum urea, creatinine as well as CRP and TNF-α compared to AZA group. Conclusions: Pumpkin seeds oil has the ability to ameliorate the biochemical pathways of the side-effects of AZA on liver and kidney.