Date of Award


Degree Name

Doctor of Philosophy



First Advisor

Dr. Susan R. Stapleton

Second Advisor

Dr. David S. Reinhold

Third Advisor

Dr. Pamela Hoppe

Fourth Advisor

Dr. Michael J. Barcelona


Type 2 diabetes is mediated by insulin resistance, the inability of insulin to elicit a normal biological response in insulin responsive tissues. Several cellular models have been utilized to determine the mechanism of induction of insulin resistance but questions remain unanswered. One model, implicates the products of the Hexosamine Biosynthetic Pathway (HBP) in the induction of insulin resistance under hyperglycemia. The major end product of HBP, UDP-GlcNAc, is the substrate for O-GlcNAc transferase, an enzyme that catalyzes the O-GlcNAcylation of numerous proteins. This modification may play a role in induction of insulin resistance and thus needs to be evaluated in different cell types. Therefore, we set out to test whether or not insulin resistance could be established in primary rat hepatocytes. To accomplish this we used a HBP precursor, glucosamine and first assessed whether or not insulin resistance was established by evaluating insulin's effect on a key signaling protein, Akt.

Results indicate that the insulin induced phosphorylation of Akt was decreased in the presence of glucosamine when compared to the control, suggesting an insulin resistant state. Signal protein activation is very important for the insulin regulation of gene expression. If the activation of signal proteins is altered, then the effect on gene expression should also be altered. Results show that under glucosamine treatment, insulin was no longer able to control the gene expression of a number of key enzymes in major metabolic pathways. Additionally an increase in O-GlcNAc modified proteins under glucosamine compared to the control was observed and a number of these proteins were identified through LC-MS.

Lastly, the effect of selenium, an insulin mimetic agent, was examined in this model system. The effects of selenium on the phosphorylation of the insulin signaling protein, Akt and on the expression of the key enzymes of the major metabolic pathways both under normal and insulin resistant conditions were examined. Results show that selenium is a potent insulin mimetic not only under the normal/control condition, but also under the insulin resistant condition as well.

Access Setting

Dissertation-Open Access