We previously used isolated adult rat hepatocyte cultures to study the ability of glucose to induce several hepatic mRNAs. However, we found that the optimal insulin concentration required to obtain the glucose effect was > 10,000 μU/ml. To test the hypothesis that the requirement for high concentrations of insulin in the culture was due to rapid loss of insulin in hepatocyte cultures, serial measurements of insulin were made at different media insulin concentrations (0–500,000 μU/ml) and glucose concentrations (5.5 and 2.75 mM). In addition, a dose-response relationship was established between media insulin concentrations and the pattern of mRNAs present in the hepatocytes determined by two-dimensional gel electrophoresis of in vitro translation products. We found that at low insulin concentrations (1000 μU/ml), 80% of the insulin was lost to the glassware, whereas at high initial insulin concentrations, ∼23% of the insulin was lost to the glassware. Placement of media into the hepatocyte culture led to further insulin disappearance with a half-life for insulin of 41.5 h at 10,000 μU/ml and 13.8 h at 100 μU/ml. We found 16 mRNAs were altered by insulin at 5.5 mM glucose and 9 mRNAs were changed by insulin at 27.5 mM glucose. After taking into consideration the distributional and metabolic losses of insulin, all but one mRNA responded to insulin within the physiologic range of portal insulin (<1–94 μU/ml). Our data indicate that the hepatocyte culture is an excellent model to study the physiologic effects of insulin on hepatic gene expression.
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Original Articles|
August 01 1987
Effects of Insulin and Glucose on Cultured Rat Hepatocyte Gene Expression
Arshag D Mooradian;
Arshag D Mooradian
Division of Endocrinology, Department of Medicine, University of Minnesota
Minneapolis, Minnesota
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Cary N Mariash
Cary N Mariash
Division of Endocrinology, Department of Medicine, University of Minnesota
Minneapolis, Minnesota
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Address correspondence and reprint requests to Cary N. Mariash, MD, Division of Endocrinology, Box 91 Mayo, University of Minnesota Hospital, Minneapolis, MN 55455.
Address for A.D.M.: Veterans Administration Medical Center, 16111 Plummer Street, Sepulveda, CA 91343.
Diabetes 1987;36(8):938–943
Article history
Received:
July 28 1986
Revision Received:
February 12 1987
Accepted:
February 12 1987
PubMed:
3297888
Citation
Arshag D Mooradian, Cary N Mariash; Effects of Insulin and Glucose on Cultured Rat Hepatocyte Gene Expression. Diabetes 1 August 1987; 36 (8): 938–943. https://doi.org/10.2337/diab.36.8.938
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