This study evaluated the role of insulin, glucagon and the epidermal growth factor (EGF) on liver regeneration after partial hepatectomy. Male Wistar rats, weighing approximately 200 g, were used. A partial hepatectomy, with resection of the medial and left lateral lobes (67.31%), was performed on the control group and seven hormone-treated groups: insulin, glucagon, EGF, insulin plus glucagon, insulin plus EGF, glucagon plus EGF, and a combination of the three hormones. The hormones were administered subcutaneously two days prior to the partial hepatectomy. The groups administered insulin were allowed to drink 20% glucose in water. Another group of rats received simulated operations, i.e., only a laparotomy was performed. The rats were killed at six, 24, 48 and 72 hours after the operation. Remnant liver weight, deoxyribonucleic acid (DNA) content, rate of DNA synthesis, mitotic index, blood glucose and serum insulin levels were measured. The results showed that: 1) the effects of single hormone administration on posthepatectomy liver regeneration were not obvious; 2) combined administration of insulin and glucagon increased the weight of the remnant liver, the DNA content, and the rate of DNA synthesis; 3) the combined administration of insulin, glucagon, and EGF increased the regeneration based on the remnant liver weight and mitotic index; and 4) there was no concordance between the change in blood glucose levels and the effect of hormones during liver regeneration.