Role for AMP-activated protein kinase in glucose-stimulated insulin secretion and preproinsulin gene expression

• DOI: 10.1042/bj20021812
• Publication Date: 2003-04-25T13:11:10Z
• Authors: Gabriela da SILVA...; Isabelle LECLERC; Aniko VARADI; Takashi TSUBOI; S. Kelly MOULE; Guy A. RUTTER
• Keywords: 0303 health sciences; Blotting, Western; Apoptosis; Cyclic AMP-Dependent Protein Kinases; 576; Cell Line; 03 medical and health sciences; Glucose; Gene Expression Regulation; Insulin Secretion; Insulin; Calcium; Phosphorylation; Protein Precursors; Proinsulin

AMP-activated protein kinase (AMPK) has recently been implicated in the control of preproinsulin gene expression in pancreatic islet β-cells [da Silva Xavier, Leclerc, Salt, Doiron, Hardie, Kahn and Rutter (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 4023–4028]. Using pharmacological and molecular strategies to regulate AMPK activity in rat islets and clonal MIN6 β-cells, we show here that the effects of AMPK are exerted largely upstream of insulin release. Thus forced increases in AMPK activity achieved pharmacologically with 5-amino-4-imidazolecarboxamide riboside (AICAR), or by adenoviral overexpression of a truncated, constitutively active form of the enzyme (AMPKα1.T172D), blocked glucose-stimulated insulin secretion. In MIN6 cells, activation of AMPK suppressed glucose metabolism, as assessed by changes in total, cytosolic or mitochondrial [ATP] and NAD(P)H, and reduced increases in intracellular [Ca2+] caused by either glucose or tolbutamide. By contrast, inactivation of AMPK by expression of a dominant-negative form of the enzyme mutated in the catalytic site (AMPKα1.D157A) did not affect glucose-stimulated increases in [ATP], NAD(P)H or intracellular [Ca2+], but led to the unregulated release of insulin. These results indicate that inhibition of AMPK by glucose is essential for the activation of insulin secretion by the sugar, and may contribute to the transcriptional stimulation of the preproinsulin gene. Modulation of AMPK activity in the β-cell may thus represent a novel therapeutic strategy for the treatment of type 2 diabetes mellitus.