Atherosclerosis is a multifactorial disease that develops and progresses in the arterial wall in response to a variety of stimuli. Among various other stimuli, hyperlipidemia is an extremely important factor that is correlated with the development of atherosclerosis. Lemon and citrus fruits contain various bioactive flavonoids, such as eriocitrin, that prevent obesity and related metabolic diseases. Therefore we concentrated on eriocitrin, a potent flavonoid with numerous therapeutic properties, particularly its beneficial lipid-lowering action in rats subjected to high fat diet. The anti-atherosclerotic efficacy of eriocitrin was assessed in rats administered a diet rich in fat. Wistar rats were divided into five groups consisting of six animals in all groups. Group I served the control, Group II was fed a high-fat diet (HFD), and the third and fourth groups were fed an HFD supplemented with varying doses of eriocitrin, and the last group was administered simvastatin for the last 30 days. Body weight, organ weight, lipid and lipoprotein parameters, cardiac and inflammatory markers, and histological examination were evaluated in animals induced with an HFD. Eriocitrin displayed a significant anti-atherosclerotic action by lowering the body weight, organ weight, reduction in lipid content, cardiac and inflammatory markers, myocardial changes confirmed by histopathology, malondialdehyde and increased antioxidant enzyme activities, nitric oxide, as well as 6-keto-PGF1α and high-density lipoprotein levels in rats fed an HFD. The findings of the experiment suggest that the anti-atherosclerotic action of eriocitrin was due to its modulatory activity in lipid metabolism. Considering the overall results of the study it can be validated that a use of flavonoid eriocitrin might be beneficial in altering HFD-induced alterations in atherosclerotic rats.