Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118.
Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118.
J Biol Chem. 2011 Dec 9;286(49):42534-42544. doi: 10.1074/jbc.M111.242412. Epub 2011 Aug 21.
Recent studies have shown that autophagy is essential for proper β-cell function and survival. However, it is yet unclear under what pathogenic conditions autophagy is inhibited in β-cells. Here, we report that long term exposure to fatty acids and glucose block autophagic flux in β-cells, contributing to their toxic effect. INS1 cells expressing GFP-LC3 (an autophagosome marker) were treated with 0.4 mm palmitate, 0.4 mm oleate, and various concentrations of glucose for 22 h. Kinetics of the effect of fatty acids on autophagy showed a biphasic response. During the second phase of autophagy, the size of autophagosomes and the content of autophagosome substrates (GFP-LC3, p62) and endogenous LC3 was increased. During the same phase, fatty acids suppressed autophagic degradation of long lived protein in both INS1 cells and islets. In INS1 cells, palmitate induced a 3-fold decrease in the number and the acidity of Acidic Vesicular Organelles. This decrease was associated with a suppression of hydrolase activity, suppression of endocytosis, and suppression of oxidative phosphorylation. The combination of fatty acids with glucose synergistically suppressed autophagic turnover, concomitantly suppressing insulin secretion. Rapamycin treatment resulted in partial reversal of the inhibition of autophagic flux, the inhibition of insulin secretion, and the increase in cell death. Our results indicate that excess nutrient could impair autophagy in the long term, hence contributing to nutrient-induced β-cell dysfunction. This may provide a novel mechanism that connects diet-induced obesity and diabetes.
最近的研究表明,自噬对于β细胞的正常功能和存活是必不可少的。然而,目前尚不清楚在什么病理条件下β细胞中的自噬会被抑制。在这里,我们报告长期暴露于脂肪酸和葡萄糖会阻断β细胞中的自噬通量,从而导致其毒性作用。用 0.4mm 棕榈酸、0.4mm 油酸和不同浓度的葡萄糖处理表达 GFP-LC3(自噬体标志物)的 INS1 细胞 22 小时。脂肪酸对自噬的影响动力学显示出双相反应。在自噬的第二阶段,自噬体的大小以及自噬体底物(GFP-LC3、p62)和内源性 LC3 的含量增加。在同一阶段,脂肪酸抑制了 INS1 细胞和胰岛中长寿命蛋白的自噬降解。在 INS1 细胞中,棕榈酸诱导酸性囊泡细胞器的数量和酸度减少了 3 倍。这种减少与水解酶活性的抑制、内吞作用的抑制和氧化磷酸化的抑制有关。脂肪酸与葡萄糖的联合作用协同抑制自噬周转率,同时抑制胰岛素分泌。雷帕霉素处理导致自噬通量的抑制、胰岛素分泌的抑制和细胞死亡的增加部分逆转。我们的结果表明,过量的营养物质可能会长期损害自噬,从而导致营养诱导的β细胞功能障碍。这可能为饮食诱导的肥胖和糖尿病提供了一种新的机制。