Introduction: The administration of HGF during intestinal adaptation is known to enhance intestinal adaptation. Glucagon has been implicated as a potential mediator of intestinal adaptation. Previous studies have shown that HGF and glucagon synergistically increase the proliferation of hepatocytes. This study was designed to determine if HGF stimulation influenced glucagon expression in the small intestine during intestinal adaptation. Methods: Adult male Sprague-Dawley rats were randomized to either a 70% massive small bowel resection group (MSBR) or an HGF treated MSBR group (MSBR-HGF). Seven days after surgery HGF was administered intravenously at 150ug/kg/day for 14 days. At day 21 ileal and jejunal mucosa was harvested. The RAE 230A GeneChip and MAS5 software were used to determine alterations in gene expression in the small intestine mucosa. Glucagon immunoreactivity was semiquantified in the ileum and jejunum using rabbit antiglucagon antibody. Results: The MSBR-HGF group had significantly greater protein and DNA content (p < 0.05) than the MSBR group. However, glucagon gene expression in the MSBR-HGF group was decreased. Immunohistostaining for glucagon in the ileum revealed no difference in intensity between the two groups. However, the MSBR-HGF group demonstrated significantly greater jejunal glucagon immunoreactivity. Conclusions: Our data suggest that the HGF induced increase in glucagon availability is disassociated from glucagon gene up-regulation. Thus, HGF may not only enhance intestinal adaptation directly, but also indirectly by increasing the “local” availability of other growth factors in the absence of their gene up-regulation.