High glucose induces dysfunction in insulin secretory cells by different pathways: a proteomic approach.

Chronic hyperglycemia is a hallmark of type 2 diabetes and can contribute to progressive beta cell dysfunction and death. The aim of the present study was to identify pathways mediating high glucose-induced beta cell demise by a proteomic approach. INS-1E cells were exposed to 25 mM glucose for a sustained period of 24 h. Protein profiling of INS-1E cells was done by two-dimensional difference gel electrophoresis, covering the pH ranges 4-7 and 6-9 (n = 4). Differentially expressed proteins (P < 0.05) were identified by MALDI-TOF/TOF and proteomic results were confirmed by functional assays. High glucose levels impaired glucose-stimulated insulin secretion and decreased insulin content. 2D-DIGE analysis revealed 100 differentially expressed proteins that were involved in different pathways. Chaperone proteins were down-regulated, protein biosynthesis and ubiquitin-related proteasomal degradation were attenuated and perturbations in intracellular trafficking and vesicle transport and secretion could be observed. Moreover, several pathways were confirmed by functional assays and a direct role for eEF2 in insulin biosynthesis was demonstrated. The present findings provide new insights in glucotoxicity and identify key target proteins for the prevention and treatment of beta cell dysfunction in type 2 diabetes.