Research Institute of Endocrinology, Medical Science Research Institute, Kyung Hee University Hospital, Seoul 130–702, Republic of Korea.
Biol Pharm Bull. 2012;35(1):121-6. doi: 10.1248/bpb.35.121.
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.
葡萄糖毒性导致β细胞进行性衰竭和显性糖尿病的发生。氧化应激是β细胞葡萄糖毒性的一个重要方面。我们研究了类黄酮芹菜素是否能保护胰岛β细胞免受 2-脱氧-D-核糖（dRib）诱导的氧化细胞损伤。在存在 dRib 的情况下，将 HIT-T15 胰岛β细胞与或不与芹菜素一起培养。使用细胞计数试剂盒（CCK-8）监测时间和剂量依赖性细胞活力，使用细胞死亡酶联免疫吸附测定（ELISA）试剂盒分析细胞凋亡。使用 JC-1 试剂盒测定线粒体膜电位（ΔΨ（m））。通过使用 2'，7'-二氯荧光素二乙酸酯（DCFH-DA）作为探针荧光分析测定细胞内氧化应激。此外，分析与氧化应激相关的转录因子核因子-κB（NF-κB）和激活蛋白 1（AP-1）的 DNA 结合活性。dRib 降低细胞存活率和ΔΨ（m），同时显著增加细胞内活性氧（ROS）水平、细胞凋亡和与氧化应激相关的转录因子 NF-κB 和 AP-1 的活性。然而，用芹菜素预处理细胞可减轻 dRib 诱导的所有作用。抗氧化剂 N-乙酰-L-半胱氨酸（NAC）和α-硫辛酸（ALA）也可防止 dRib 诱导的氧化损伤和 NF-κB 和 AP-1 的激活。总之，这些结果表明，芹菜素通过氧化应激相关信号减轻 dRib 诱导的胰岛β细胞损伤。
