Apigenin Attenuates 2-Deoxy-D-ribose-Induced Oxidative Cell Damage in HIT-T15 Pancreatic β-Cells

Glucose toxicity contributes to progressive beta-cell failure and the development of overt diabetes. Oxidative stress is an important aspect of glucose toxicity in pancreatic beta-cells. We investigated whether the flavonoid apigenin protects pancreatic beta-cells from 2-deoxy-D-ribose (dRib)-induced oxidative cell damage. HIT-T15 pancreatic beta-cells were cultured with or without apigenin in the presence of dRib. Time- and dose-dependent cell viability was monitored using a cell counting kit (CCK-8), while the induction of apoptosis was analyzed using a cell death enzyme-linked immunosorbent assay (ELISA) kit. Mitochondria] membrane potential (Delta Psi(m),) was determined using the JC-1 kit. Intracellular oxidative stress was measured by fluorometric analysis of DCFH oxidation using 2',7'-dichlorofluorescin diacetate (DCFH-DA) as the probe. In addition, the DNA binding activity of the oxidative stress-related transcriptional factors nuclear factor-kappa B (NF-kappa B) and activator protein 1 (AP-1) were analyzed. dRib reduced cell survival and Delta Psi(m), while it markedly increased intracellular levels of reactive oxygen species (ROS), apoptosis, and the activity of the oxidative stress-related transcription factors NF-kappa B and AP-1. However, pretreatment of cells with apigenin attenuated all the dRib-induced effects. The anti-oxidants, N-acetyl-L-cysteine (NAC) and alpha lipoic acid (ALA), also prevented both dRib-induced oxidative damage and activation of NF-kappa B and AP-1. Taken together, these results suggest that apigenin attenuates dRib-induced cell damage in pancreatic beta-cells via oxidative stress-related signaling.