An animal study was carried out to evaluate the influence of dietary fenugreek seeds on regression of preestablished cholesterol gallstones (CGS). CGS was induced by feeding a high-cholesterol diet for 10 weeks. After CGS induction, the animals were maintained for a further 10 weeks on experimental diets of high cholesterol, 6% fenugreek powder, 12% fenugreek powder, or basal control. Incidence of CGS and its severity were evaluated at the end of this feeding regimen. The incidence of CGS was significantly lowered as a result of dietary fenugreek seeds, the extent of regression being 61% and 64% in the low and high dose groups compared with 10% regression in the basal control group. The antilithogenic influence of dietary fenugreek was accompanied by significant reductions of more than 35% in serum cholesterol concentration. Hepatic cholesterol concentration was also profoundly lowered by dietary fenugreek, being 53%-63% lower than that of the basal control diet. Biliary cholesterol concentration was significantly lower as a result of dietary fenugreek during the post-CGS induction period, resulting in a decreased cholesterol:phospholipid ratio (0.44 and 0.40 compared with 0.79 in the basal control group). Biliary cholesterol : bile acid ratio was lowered by 67% and 73% upon feeding fenugreek, significantly lower than that in the basal control group. The cholesterol saturation index in the bile was also beneficially lowered by fenugreek treatment during the post-CGS induction period (the index was 0.90 and 0.42 compared with 1.86 in the basal control group). The present study provides evidence of the potency of hypolipidemic fenugreek seeds in regressing preestablished CGS, and this beneficial antilithogenic effect is attributable to its primary influence on cholesterol levels. This finding is significant in the context of evolving a dietary strategy to address CGS, which could help in preventing the incidence and regression of existing CGS and controlling possible recurrence.