Background: In the developmental stage, pancreas derives from the endodermal cells where the transcription factor, pancreatic duodenal homeobox gene-1 (pdx-1) is expressed. In adulthood, pdx-1 expression is localized to pancreatic beta cells, which is necessary for maintenance of beta cell function. Recently, ectopic expression of pdx-1 in the liver successfully induced insulin production and ameliorated hyperglycemia. Our study was designed to investigate the effects of forced expression of pdx-1 in ileal epithelia by adenovirus-mediated gene transfer.
Methods: The recombinant, replication-deficient adenovirus carrying the pdx-1 gene was constructed using the COS-TPC method. ICR mice were treated with intraperitoneal injection of 220 mg/kg streptozotocin (STZ). After determining the hyperglycemia, a loop of ileum was constructed and the adenovirus solutions (Ad-pdx-1 and Ad-lacZ 1 x 10(8) PFU/body) were injected into the lumen of the ileal loop. In this model, immunohistochemical or fluorescent analyses of PDX-1 and insulin in the adenovirus-infected ileal epithelia were carried out. Reverse transcription polymerase chain reaction of pdx-1 and other pancreatic markers were investigated. Blood glucose concentrations were measured by drawing blood from ocular veins. Immunoreactive insulin extracted from the adenovirus-infected ileum was measured.
Results: Ad-pdx-1 induced ectopic PDX-1 expression in the ileum. The PDX-1 positive cells in the ileal epithelia were positive for insulin; mRNA of insulin-1, insulin-2 and pdx-1 were expressed in mice infected with Ad-pdx-1. Hyperglycemia was improved in STZ-treated mice infected with Ad-pdx-1. Immunoreactive insulin in the ileum extract was increased significantly in mice with Ad-pdx-1.
Conclusions: Gene transfer of PDX-1 in intestinal epithelia could be a promising strategy for diabetes mellitus by inducing ectopic insulin producing cells.