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Apigenin attenuates 2-deoxy-D-ribose-induced oxidative cell damage in HIT-T15 pancreatic β-cells
Kwang Sik Suh1, Seungjoon Oh, Jeong-Taek Woo
1Research Institute of Endocrinology, Medical Science Research Institute, Kyung Hee University Hospital, Seoul 130–702, Republic of Korea.
Abstract:
Glucose toxicity contributes to progressive β-cell failure and the development of overt diabetes. Oxidative stress is an important aspect of glucose toxicity in pancreatic β-cells. We investigated whether the flavonoid apigenin protects pancreatic β-cells from 2-deoxy-D-ribose (dRib)-induced oxidative cell damage. HIT-T15 pancreatic β-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. Mitochondrial membrane potential (ΔΨ(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-κB (NF-κB) and activator protein 1 (AP-1) were analyzed. dRib reduced cell survival and ΔΨ(m), while it markedly increased intracellular levels of reactive oxygen species (ROS), apoptosis, and the activity of the oxidative stress-related transcription factors NF-κB 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-κB and AP-1. Taken together, these results suggest that apigenin attenuates dRib-induced cell damage in pancreatic β-cells via oxidative stress-related signaling.
Insights
The flavonoid apigenin protects pancreatic beta cells from oxidative stress and damage induced by 2-deoxy-D-ribose (dRib). Apigenin treatment attenuated dRib-induced cell death, reactive oxygen species generation, and inflammatory signaling pathways.
Area of Science:
- Endocrinology
- Cell Biology
- Biochemistry
Background:
- Glucose toxicity is a key factor in pancreatic beta-cell dysfunction and diabetes development.
- Oxidative stress plays a significant role in glucose toxicity within pancreatic beta-cells.
Purpose of the Study:
- To investigate the protective effects of the flavonoid apigenin against 2-deoxy-D-ribose (dRib)-induced oxidative damage in pancreatic beta-cells.
- To elucidate the underlying mechanisms of apigenin's protective action, focusing on oxidative stress and inflammatory signaling.
Main Methods:
- HIT-T15 pancreatic beta-cells were treated with apigenin prior to dRib exposure.
- Cell viability (CCK-8), apoptosis (ELISA), mitochondrial membrane potential (JC-1), and reactive oxygen species (ROS) generation (DCFH-DA) were assessed.
- DNA binding activity of NF-κB and AP-1 was analyzed.
Main Results:
- dRib significantly reduced cell survival and mitochondrial membrane potential, while increasing ROS production, apoptosis, and NF-κB/AP-1 activity.
- Apigenin pretreatment effectively counteracted all dRib-induced detrimental effects.
- Antioxidants N-acetyl-L-cysteine (NAC) and alpha lipoic acid (ALA) demonstrated similar protective effects against dRib-induced damage.
Conclusions:
- Apigenin exhibits protective properties against dRib-induced oxidative stress and cell damage in pancreatic beta-cells.
- The protective mechanism of apigenin involves the attenuation of oxidative stress-related signaling pathways, including NF-κB and AP-1 activation.
- Apigenin represents a potential therapeutic agent for mitigating beta-cell dysfunction associated with glucose toxicity.