Cytotoxic Effect of \" Glycated Albumin-Transition Metal Ion\" on Rat Hepatocyte Suspension

Background: Combination of glycation and oxidation is associated with diabetes mellitus. The aim of this study was to clarify the effect of glycated proteins in presence of transition metal ions on production of reactive oxygen species (ROS) in rat hepatocyte suspension.  Methods: Glycated albumin was prepared by incubation of bovine serum albumin with 100 mM glucose in 0.3 M phosphate buffer at 37°C for 2, 4 , and 6 weeks. The prepared rat liver cell suspension was treated with glycated albumin in presence of either Fe+++ or Cu++ ions. Produced malone-dialdehyde was measured as an indicator of ROS and of cell injury.  Results: The results showed Fe+++ and Cu++ ions increase the ROS production in presence of glycated albumin (p<0.01). All prepared glycated albumin showed cytotoxicity in rat hepatocytes suspension in the presence of cupric and ferric ions, and this injury was dependent to metal ion concentration. Higher degree of glycation showed higher effect on ROS production (p<0.01). Comparing the effect of Fe+++ and Cu++, cupric ion had higher cytotoxic effect (p<0.01).  Conclusion: These results indicated that hepatocytes may be damaged by ROS which are produced by the interaction of the glycated albumin and transition metal ion.