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Effects of insulin and vanadate on glucose production in cultured hepatocytes isolated from rats on poly- and monounsaturated fat diet

Insulin resistance is a common phenomenon in obesity and Type 2 diabetes. Increased dietary fat intake will lead to impairment of insulin action. The aim of this study was to find out changing of hepatic glucose output in dependence of fat diet and possible direct action of vanadate and insulin on glucose production in cultured hepatocytes. Hepatocytes were isolated by a collagenase perfusion technique and cultured for 24 h in M 199 serum-free medium. The glucose production was measured by incubating the cultures in glucose-free Hanks-Hepes medium with addition 10 mmol/l pyruvate. The glucose released into the medium was determined enzymatically with glucose oxidase. The glucagon stimulated glucose production in hepatocytes isolated from rats on high polyunsaturated fat diet was significantly higher comparable with rats on high monounsaturated fat diet. Insulin significantly decreased and normalized basic glucose production in he patocytes isolated from rats on high monounsaturated fat diet. Insulin did not decrease glucagon stimulated glucose production in both experimental groups. On the other hand vanadate significantly decreased glucagon stimulated glucose production in both experimental groups. Rats on high polyunsaturated fat diet had greater increase of hepatic glucose production and lower insulin sensitivity, suggesting development of insulin resistance. Vanadate partially normalized glucagon stimulated glucose production probably inhibiting gluconeogenesis.

fat diet; glucose production; hepatocytes; isulin; vanadate

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