Exercise generally has been found to produce beneficial effects on plasma lipid and lipoprotein profiles, but the mechanisms involved and possible dietary interactions have not been well defined. Weanling male Fischer 344 rats were assigned to three groups: normocholesterolemic diet sedentary (NS), hypercholesterolemic diet sedentary (HS), and hypercholesterolemic diet exercised (HE). Exercise consisted of treadmill running at 1.2 to 1.4 km/h at a 9% grade, six days weekly, for a 10-week experimental period. Lipoproteins from plasma and from a recirculating in situ liver perfusion system were then isolated and analyzed. The values of several parameters for HE tended to fall intermediate between HS and NS. Final total plasma cholesterol and liver cholesterol concentrations were significantly different among all three groups (HS greater than HE greater than NS). Plasma HDL-cholesterol and phospholipids and perfusate HDL-cholesterol production rate per gram liver were all significantly lower in HS v NS, with HE lying in between. Plasma HDL protein was lower in HS than in both other groups. Plasma total triglyceride levels were significantly lowered by exercise, but neither plasma nor perfusate VLDL triglyceride levels differed significantly among the three groups. Food intakes of HE and HS rats were similar, but HE rats had significantly lower final body weights. The results suggest that (1) exercise may ameliorate many of the changes in lipoprotein and cholesterol metabolism induced by a diet containing lard and cholesterol, and (2) some of these changes may be mediated by changes in hepatic lipoprotein production.