Depleted uranium (DU) and cesium-137 (137Cs) are radionuclides spread in the environment due to industrial activities, incidents or accidents. This pollution sets a risk of human exposure to low levels of radiations through contaminated foodstuff. The impact of a chronic ingestion of DU or 137Cs on cholesterol metabolism in the liver and the brain has been studied. Indeed, cholesterol is crucial in physiology, being a component of cell membranes and a precursor to numerous molecules (bile acids…). Disruption of its metabolism is associated to many pathologies such as atherosclerosis or Alzheimer’s disease. Rats daily ingested a low level of DU or 137Cs over 9 months. For each radionuclide, a reference model (rats contaminated since adulthood) and a more sensitive model (hypercholesterolemic or contaminated since fetal life) were studied. The effects mainly consist of changes in gene expression or enzymatic activity of various actors of cholesterol metabolism. DU mainly affects one catabolism enzyme in both models, as well as membrane transporters and regulation factors. 137Cs mainly affects the storage enzyme in both models as well as catabolism enzymes, apolipoproteins, and regulation factors. No change in the plasma profile or in the tissue concentration of cholesterol (hepatic/cerebral) is recorded, whatever the model and the radionuclide. Thus, a chronic internal contamination with DU or 137Cs induces molecular modifications in cholesterol metabolism in the rat, without affecting its homeostasis or the general health status in all of our experimental models.