Objective The mechanisms of glucotoxicity involve several transcription factors and are, at least in part, mediated by generation of chronic oxidative stress (8). Several recent studies (6,7) supported that the Fatty Acid Translocase Cluster Determinant 36 (FAT/CD36) increases uptake of free fat acid in high glucose state in several cells. Yet, little is known about the relation between glucotoxicity and CD36 in the islets. The purposes of the present study were to determine whether prolonged exposure of pancreatic islets to glucotoxic condition disrupts CD36 or increases CD36, we also wanted to evaluate a role of CD36 in pancreatic islets against glucotoxicity. Methods We checked intracellular peroxide levels, insulin transcription factors, insulin mRNA, glucose stimulated insulin secretion (GSIS) and CD36 of INS-1 cells. INS-1 cells were transfected of CD36 siRNA for inhibition of CD36 in INS-1 cells. Results The intracellular peroxide levels of the pancreatic islets were increased in the high glucose (30 mM glucose in INS-1 cell) media compared to 5.6 mM glucose media. The insulin mRNA levels and GSIS were decreased in INS-1 cells exposed to high glucose media compared to 5.6 mM glucose media. The insulin mRNA levels, PDX-1 and GSIS were decreased in INS-1 cells exposed to glucotoxic condition (30 mM glucose) compared to 5.6 mM glucose media. And, at INS-1 cells exposed to glucotoxic condition, CD 36 was significant increased compared to normal glucose media at 12 hours. After transfection of CD36 siRNA, CD 36 was significantly decreased compared to control and insulin mRNA and GSIS were significantly increased compared to without CD36 siRNA in INS-1 cells. Conclusion These results suggest that the influx of fatty acid to islet at early high glucose condition can be increased by increasing of CD36. Therefore, at islets exposed to high glucose and high fatty acid may be exacerbating beta cell dysfunction via increasing of CD 36 and inhibition of CD36 may be prevent glucotoxicity in INS-1 cells.