The fruits of Opuntia ficus-indica (L.) Mill. (OFI), a member of the family Cactaceae, is cultivated in Saudi Arabia and considered as a functional food with anti-inflammatory activity.The aim of current study was to chemically characterise OFI growing in Saudi Arabia by GC-MS and evaluate its anti-inflammatory activity in rat models via carrageenan-induced paw oedema and croton oil-induced ear oedema at two doses 100 and 200 mg/kg. Palmitic acid (10.68%), linoleic acid (5.9%), oleic acid (8.16%) and β-sitosterol (24.98%) are the major constituents in OFI seed oil. In carrageenan-induced rats, the OFI seed oil (100 and 200 mg/kg) produced significant inhibition of oedema by approximately 46% and 62%, respectively, and reduced PGE2 concentrations in exudates by 54% and 67%, respectively. Also, it significantly decreased the weight of punch from challenged ears by 20% and 33%, and myeloperoxidase (MPO) activity by 54% and 62% of the induced ear respectively. This was accompanied by amelioration of croton oil induced histopathological changes. In conclusion, the antiinflammatory activity of the OFI seed oil might be attributed to the presence of USFA such as oleic acid (omega-9) in addition to β-sitosterol through decreasing PGE2 and MPO activity in the inflamed tissues which was supported by histopathological examination.