Whether glucocorticoid-responsive gluconeogenesis is an ancestral or later-derived feature of vertebrates is unknown. The purpose of this study is to investigate carbohydrate metabolism of the liver in response to generalized stress in Pacific hagfish (Eptatretus stoutii). Pacific hagfish were exposed to an acute (30 min) bout of generalized stress and sampled for blood and liver at different time points during recovery (30 min, 6 h, 24 h, and 72 h). Within 6 h post-stress, blood glucose levels increased from 1.4 ± 0.1 mM to 3.3 ± 0.3 mM. Likewise, blood lactate levels increased from 3.1 ± 0.4 mM to 5.5 ± 1.3 mM. The transcriptional regulation of genes which participate in carbohydrate metabolism in the liver were examined. These findings suggest that carbohydrate metabolism is responsive to stress in hagfish, and further examination of the metabolism of the liver may shed light on the biochemical pathways involved in these responses. Our understanding of the metabolic physiology in this ancient lineage of vertebrates can contribute to our fundamental understanding of how vertebrate processes of carbohydrate handling evolved.