Abstract In vitro physicochemical and antioxidant properties of mixture of Flax/Sesame (LS) and Flax/Peanut (LA) and in vivo hypolipidemic, antioxidant and hepatoprotective activities were carried out to ascertain the claim of its utilisation against diseases. The seeds mixture rich in unsaturated fatty acids were prepared with 5/1 ratio of ω-6/ω-3 fatty acids and were orally administered ad libitum to rats by standard diet for 30 days. High cholesterol fed diet rats (CD-chol) exhibited a significant increase in total plasma and liver lipid parameters and atherogenicity and a significant decrease in high-density lipoproteins (HDL) and HDL/TC ratio (HTR). Administration of (LS) or (LA) seeds mixture to hypercholesterolemic rats (MS-LSchol and MS-LAchol groups respectively) significantly ameliorated lipid parameters and showed an increase of PUFAs (ALA and LA) and MUFAs and a decrease of SFAs in plasma and liver of MS-LSchol and MS-LAchol groups. Furthermore, malondialdehyde levels decreased and the efficiency of antioxidant defense system was improved compared to CD-chol group. Liver histological sections showed lipid storage in hepatocytes of CD-chol group and an improvement was noted in both supplemented groups. Our results suggested that seeds mixtures of Flax/Sesame and Flax/Peanut have anti-atherogenic and hepatoprotective effects.