In this study, effect of digoxigenin-3-O-rutin isolated from the seeds of Trigonella foenum graecum on thyroid hormones and serum lipid concentrations was evaluated in L-thyroxine (L-T 4 )-induced hyperthyroidism in rats. Digoxigenin-3-O-rutin was administered (10mg/kg) to L-thyroxine (L-T 4 )-induced hyperthyroidic rats and alterations in the concentrations of serum thyroid hormones, insulin, glucose, hepatic 5′-monodeiodinase (5′DI) and glucose-6-phosphatase (G-6-Pase) activity were analyzed. Antioxidant status was estimated by determining the levels of antioxidative enzymes and lipidperoxidation. L-T 4 (500μg/kg, s.c./d) administration increased the serum levels of thyroxine (T4), triidothyronine (T3), glucose, insulin, different lipids, activity of hepatic 5′-DI and G-6-Pase. High lipidperoxidation level was observed both in liver and cardiac tissues with a depletion in cellular antioxidants. On the contrary, test drug (10mg/kg) treatment improved the alterations with respect to hormonal levels, lipid concentrations and lipid peroxidation towards normalcy and enhanced the antioxidant activities. Rats treated with PTU generally gave lower results compared to groups treated with the test drug. The antithyroidic role of the test compound is mediated possibly through the inhibition in 5′DI activity. Improvement in lipid profile by the test drug might have protective effect on cardiovascular health in vivo.