Department of Food and Nutrition, College of Medical and Life Science, Silla University, Busan 617-736, Korea.
Department of Food Science and Nutrition, Pusan National University, 2, Busandaehak-ro 63beon-gil, Geumjeong-gu, Busan 609-735, Korea.
Nutr Res Pract. 2014 Oct;8(5):494-500. doi: 10.4162/nrp.2014.8.5.494. Epub 2014 Jun 30.
BACKGROUND/OBJECTIVES: This study investigated whether Padina arborescens extract (PAE) protects INS-1 pancreatic β cells against glucotoxicity-induced apoptosis.
MATERIALS/METHODS: Assays, including cell viability, lipid peroxidation, generation of intracellular ROS, NO production, antioxidant enzyme activity and insulin secretion, were conducted. The expressions of Bax, Bcl-2, and caspase-3 proteins in INS-1 cells were evaluated by western blot analysis, and apoptosis/necrosis induced by high glucose was determined by analysis of FITC-Annexin V/PI staining.
Treatment with high concentrations of glucose induced INS-1 cell death, but PAE at concentrations of 25, 50 or 100 µg/ml significantly increased cell viability. The treatment with PAE dose dependently reduced the lipid peroxidation and increased the activities of antioxidant enzymes reduced by 30 mM glucose, while intracellular ROS levels increased under conditions of 30 mM glucose. PAE treatment improved the secretory responsiveness following stimulation with glucose. The results also demonstrated that glucotoxicity-induced apoptosis is associated with modulation of the Bax/Bcl-2 ratio. When INS-1 cells were stained with Annexin V/PI, we found that PAE reduced apoptosis by glucotoxicity.
In conclusion, the present study indicates that PAE protects against high glucose-induced apoptosis in pancreatic β cells by reducing oxidative stress.
背景/目的:本研究旨在探讨鹿角菜提取物(PAE)是否能保护 INS-1 胰腺 β 细胞免受糖毒性诱导的细胞凋亡。
材料/方法:进行了细胞活力、脂质过氧化、细胞内 ROS 生成、NO 产生、抗氧化酶活性和胰岛素分泌的测定。通过 Western blot 分析评估 INS-1 细胞中 Bax、Bcl-2 和 caspase-3 蛋白的表达,通过分析 FITC-Annexin V/PI 染色确定高葡萄糖诱导的细胞凋亡/坏死。
高浓度葡萄糖处理诱导 INS-1 细胞死亡,但 25、50 或 100μg/ml 的 PAE 显著增加细胞活力。PAE 处理剂量依赖性地降低了由 30mM 葡萄糖引起的脂质过氧化,并增加了被 30mM 葡萄糖降低的抗氧化酶活性,而在 30mM 葡萄糖条件下,细胞内 ROS 水平升高。PAE 处理改善了葡萄糖刺激后的分泌反应性。结果还表明,糖毒性诱导的细胞凋亡与 Bax/Bcl-2 比值的调节有关。当用 Annexin V/PI 对 INS-1 细胞进行染色时,我们发现 PAE 减少了由糖毒性引起的细胞凋亡。
综上所述,本研究表明 PAE 通过降低氧化应激来保护胰腺 β 细胞免受高葡萄糖诱导的细胞凋亡。