Chronic elevated glucose is harmful to pancreatic beta-cells, resulting in pancreatic beta-cells dysfunction and apoptosis. Understanding the molecular mechanisms associated with beta-cells survival is pivotal for the prevention of beta-cells injury caused by glucotoxicity. The role of Phosphatase and tensin homologue (PTEN)-induced putative kinase 1 (PINK1) in the fate of pancreatic beta-cells constantly exposed to high glucose was studied. Sustained high glucose increased PINK1 protein expression both in rat pancreatic beta-cells and INS-1 beta-cells, and that this increase can be inhibited by PINK1 knockdown and further enhanced by PINK1 over-expression. PINK1 deficiency aggravated glucotoxicity-induced pancreatic beta-cells apoptosis and inhibition of autophagy whereas PINK1 could reverse these adverse effects. This study provides fundamental data supporting the potential protective role of PINK1 as a new therapeutic target necessary to preserve beta-cells survival under non-physiological hyperglycemia conditions. (C) 2016 Elsevier Inc. All rights reserved.