The effects of insulin, glucagon, isoproterenol and carbachol on the regeneration of injured liver were investigated in rats treated with carbon tetrachloride (CCl4). These agents effectively potentiated hepatic DNA synthesis in rats both at 48 and at 72 hr after CCl4 intoxication. The maximal stimulatory effects of the agents on the synthesis coincided in time with the peak of elevation in basal DNA synthesis following the intoxication. Plasma levels of insulin and triiodothyronine were decreased before the elevation of basal DNA synthesis in CCl4-treated rats. The possible relationship of these changes in plasma hormones to the potentiated effects of the agents on DNA synthesis was examined in rats treated with streptozotocin (STZ) or methylthiouracil (MTU). The agents caused no potentiation in STZ-treated rats. On the other hand, in MTU-treated rats, isoproterenol and carbachol significantly stimulated DNA synthesis, but this was not the case with insulin and glucagon. These results suggest that the pancreatic hormonal, beta-adrenergic and cholinergic stimulations play positive roles in regulating liver regeneration after CCl4 intoxication. Furthermore, the hypothyroid state developed in CCl4-treated rats may provide favorable conditions for the stimulation of DNA synthesis by isoproterenol and carbachol. It is unlikely, however, that insulin deficiency contributes to potentiations in the regenerative responses of the injured liver.