A Cellular Assay for Measuring the Modulation of Glucose Production in H4IIE Cells
2006; Mary Ann Liebert, Inc.; Volume: 4; Issue: 5 Linguagem: Inglês
10.1089/adt.2006.4.525
ISSN1557-8127
AutoresAnne-Marie de Raemy-Schenk, Solenne Troublé, P. Gaillard, Patrick Page, Jean‐Pierre Gotteland, Alexander Scheer, Paul Lang, Karen Yeow,
Tópico(s)Protein Kinase Regulation and GTPase Signaling
ResumoType II diabetes and its associated complications are a major health concern of the developed world. One of the hallmarks of diabetes is insulin resistance, where secreted insulin no longer has any effect on its target tissues, namely, liver, muscle, and fat. An important therapeutic strategy is to modulate blood glucose levels using pharmacological agents. Glycogen synthase kinase-3 (GSK3) is a serine-threonine protein kinase that plays important roles in regulating glucose metabolism. It is a key negative regulator of insulin action and is an important contributing factor to insulin resistance in liver, muscle, and adipose tissue. We describe the development of a cell-based assay designed to measure glucose production in rat hepatoma cell line H4IIE liver cells in response to treatment with small molecule inhibitors, including GSK3 inhibitors. The assay is set up in a 96-well format, and glucose production is assessed using a convenient fluorescence-based readout. This disease-relevant cellular assay is a valuable tool for the progression of small molecules that modulate glucose production.
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