OBJECTIVE To evaluate the splenic uptake function after irradiation with high-energy X-rays. METHODS Fourteen male Wistar rats were distributed into three groups. Group 1 (n = 6) - control, non-irradiated; Group 2 (n = 4) - animals that were irradiated and studied 24 h after irradiation; and Group 3 (n = 4) - animals that were irradiated and studied 48 h after irradiation. The animals were irradiated with 8 Gy X-rays in the abdominal region. According with the groups, after 24 or 48 h, 1 ml/kg of a 50% colloidal carbon solution was injected in the left internal jugular vein. After 40 min, the spleens were removed for histological studies. Macrophages containing carbon pigments in their cytoplasms were counted in 16 consecutive microscopic fields, and their means were considered as the uptake pattern of each animal. RESULTS In the control groups, carbon pigments were captured by macrophages in the red and white pulps, while in the irradiated groups, the uptake in the marginal zone, around the white pulp, was enhanced. There was no disorder on the splenic parenchyma or necrosis in histological analyzes. Qualitatively rare apoptotic events were observed, with no difference between control and irradiated animals. CONCLUSIONS The high-energy X-ray, used in radiotherapy, modifies the splenic clearance, enhancing the amount of marginal zone macrophages containing colloid particles. This radiation was not associated with morphological changes, nor with necrosis or apoptosis of splenic tissue.