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(Objective)The periparturient cows undergoes an experience from pregnancy to lactation,and increased demand for glucose to support milk production,which will result in a negative energy balance (NEB)with hypoglycemia.However,hormonal regulation of the balance of blood glucose is of critical importance in maintaining the metabolic equilibrium for lactating dairy cows.But,the effect of glucagon(GLN) on gluconeogenesis in dairy cows was equivocal.So,the objective of the present study was to describe the effect of GLN on hepatic gluconeogenesis and nutrient metabolism in dairy cows.(Method)The expression and activity of pyruvate carboxylase (PC) and phosphoenolpyruvate carboxykinase(PEPCK) were evaluated in vitro cultured calf hepatocytes treated with various concentrations of GLN by real-time PCR and spectrophotometry,respectively.(Result) The results showed that mRNA levels and enzyme activity of PC and PEPCK in hepatocytes increased dose dependently with increasing GLN concentration.(Conclusion)These results indicate that high concentration GLN promotes hepatocyte gluconeogenesis,which may improve NEB and futher offer the basis for preventing the NEB-related metabolic disorders in dairy cows.