To study the effects of the subcutaneous injection of glucagon and/or oral administration of glycerol on blood metabolites and hormones of Holstein dairy cows induced with fatty liver disease, twenty multiparous cows were fed a dry cow ration supplemented with 12 kg of cracked corn during the dry period to increase the likelihood of fatty liver disease development. Cows with a body condition score (BCS) of ≥ 3.5 points (0-5 scale) were randomly assigned to one of four treatment groups--saline, glucagon, glucagon plus glycerol, and glycerol. Following treatment, serial blood samples were collected to determine the effect of glucagon and/or glycerol on blood composition. Glucagon injection alone increased postpartal plasma glucose, glucagon, and insulin and decreased plasma NEFA and TAG. Glucagon plus glycerol treatment increased and sustained postpartal plasma glucose and insulin and decreased postpartal plasma NEFA and TAG. Administration of glycerol alone increased plasma glucose and decreased plasma NEFA during the postpartal period. Early postpartal treatment of cows with glucagon and/or glycerol increased plasma glucose and decreased plasma NEFA. This response would suggest that these treatments would decrease the likelihood of fatty liver disease in dairy cows.