| This dissertation describes the effect of glucose intake on MNC in vivo and monocytic cell lines in vitro. It describes the effect of glucose on the activity of transcription factors and the expression of genes known to affect the integrity of the vasculature. The electromobility shift assay has been utilized to demonstrate that the ingestion of glucose activates NF-kappaB. Western blots and an activity assay have been utilized to demonstrate that protein levels and the activity of certain components of the NF-kappaB signaling pathway are altered. Real time polymerise chain reaction (PCR) and the Atlas(TM) Human Cardiovascular cDNA Expression Array were used to identify some of the genes modulated by glucose at the transcriptional level. Some of these genes that were identified are known to affect the integrity of the cells of the vasculature or cells of the immune system when exposed to high levels of glucose or in the diabetic state. These genes include tumor necrosis factor alpha (TNFalpha), Integrin beta1, Integrin beta2, CD69, and intercellular adhesion molecule-3 (ICAM-3) mRNA. The SuperArray(TM) GEArray Q Series Human Insulin Signaling Pathway Gene Array has been used to determine whether ingestion of glucose alters mRNA levels of key genes along the insulin signaling pathway.; Our findings show that an acute increase in blood glucose levels achieved through ingestion of 75 grams of glucose alters the expression of genes in human MNC that can potentially lead to detrimental effects in the vasculature or to an alteration in the expression of genes along the insulin signaling pathway. These effects depend upon the ability of glucose to activate a variety of transcription factors that can then regulate gene expression at the transcriptional level. One specific example demonstrated in this dissertation is the activation of the transcription factor NF-kappaB, which is due to the activation of components of the NF-kappaB signaling pathway in MNC. Ultimately, the increase in mRNA expression of TNFalpha may be due to the activation of NF-kappaB. The mRNA expression of TNFalpha, Integrin beta1, Integrin beta2, CD69, and ICAM-3 increase in human MNC one and two hours following a 75 gram glucose challenge.{09}Glucose alters the expression of many genes along the insulin signaling pathway. (Abstract shortened by UMI.)... |
