Objective: To establish INS-1 cell iron overload model, and study the effect on proliferation, insulin-secretion and oxidative stress change.
Methods: INS-1 cells were cultured in either normal or high glucose medium. Both groups were treated with different concentrations of (5, 10, 20, 40, 80 μmol/l respectively) of ferric ammonium citrate (FAC). Number and state of the cells were detected by trypan blue staining. Labile iron pool (LIP) were calculated by detecting calcein-AM fluorescence. Insulin levels were detected using ELISA kit. After establishment of iron overload model, related oxidative stress (ROS) levels were detected in both normal and high glucose groups.
Results: 1) there were no significant differences of INS-1 cell numbers among different FAC concentration groups and control group after 24h or 48h. LIP level significantly increased with the FAC concentration. 2) In normal glucose medium groups, insulin levels had no correlation with FAC concentration. While in high glucose medium groups, insulin levels were first increased then dropped down with increase of FAC concentration. 3) In iron overload model, ROS level significantly increased in both normal and high glucose groups when compared with control.
Conclusions: To a certain concentration, FAC did not affect INS-1 cell proliferation. Co-cultured in high glucose medium, iron overload may cause glucose metabolic disorder in INS-1 cells, resulting in insulin resistance as well as decrease of insulin secretion. Iron overload can induce significant increase of ROS in INS-1 cells, which is possibly related to the change of cell function and insulin secretion.
27 - 29 Sep 2018
European Society for Paediatric Endocrinology