ANTI-INFLAMMATORY STRATEGIES TO PROTACT PANCREATIC β-CELSS FUNCTION IN A CELLULAR MODEL OF TYPE 2 DIABET

Abstract

Diabetes mellitus is initiated by defective or reduced circulating insulin due to an incapability to transmit glucose from the bloodstream into the tissues where it is used for energy. The beta (β) cells are the main cell type in the islet, which act to release the insulin and comprise more than 80% of the islet mass. Β-cell exposure to high levels of glucose and fatty acids, glucolipotoxicity, can also stimulate inflammatory pathways, impairing cell survival and function. This study investigated the response of INS-1 cells to high levels of glucose and fatty acids. The cells were treated with control (RPMI-1640 including 11 mM glucose), GLT RPMI-1640 (27 mM glucose, 200 μM oleic acid, 200 μM palmitic acid) and GLT plus carnosine RPMI-1640 (27 mM glucose, 200 μM oleic acid, 200 μM palmitic acid and 10 mM carnosine), lysed and the extracted proteins separated by SDS gel electrophoresis. After transfer to the nitrocellulose membrane, immunoblotting was performed using NF-κB p65 and phosphorylated NF-κB p65 antibodies, normalising the data to alpha tubulin. The blots were imaged using the GeneGnome XRQ System, showing that GLT induced NF-κB p65 activation less in comparison to the control, indicating no inflammation or impairment of β-cells. In addition, there was evidence of a difficulty in the phosphorylation to regulate gene transcription of the NF-κB p65 pathway and increased inflammation of pancreatic β-cells as a result of elevated expression of the phosphorylated NF-κB p65 antibody in GLT conditions. Carnosine induced a slight overexpression of NF-κB p65 and NF- κB p65 phosphorylation, indicating that carnosine can inhibit the GLT induced upregulation of NF-κB p65 and phosphorylated NF-κB p65, thereby protecting the β-cells against nitrosative stress and providing antioxidant protection.