Impaired pancreatic duct secretion is frequently observed in insulin-dependent diabetes mellitus (IDDM), although the cellular mechanism(s) of dysfunction remains unknown. Studies in other tissues have suggested that a hyperglycemia-induced decrease in Na,K-ATPase activity could contribute to the metabolic complications of IDDM and that increased polyol metabolism is involved in this response. The present studies examined the effects of glucose on Na,KATPase activity and on expression and activity of aldose reductase (AR), a primary enzyme of polyol metabolism, in Capan-1 human pancreatic duct cells. Increasing medium glucose from 5.5 to 22 mM caused a 29% decrease in Na,KATPase activity. The decrease was corrected by 100 M sorbinil, a specific AR inhibitor. Increasing glucose from 5.5 to 110 mM also resulted in concentration-dependent increases in AR mRNA and enzyme activity that could be resolved into two components, one that was glucose specific and observed at pathophysiological concentrations ( Ͻ 55 mM) and a second that was osmotically induced at high concentrations ( Ͼ 55 mM) and which was not glucose specific. The present study demonstrates that pathophysiological levels of glucose specifically activate polyol metabolism with a consequent decrease in Na,K-ATPase activity in pancreatic duct epithelial cells, and that this response to hyperglycemia could contribute to decreased pancreatic secretion observed in IDDM. This is the first report of AR regulation in the pancreatic duct epithelium. ( J. Clin. Invest. 1997. 100: 1685-1692.)