Liraglutide prevents high glucose level induced insulinoma cells apoptosis by targeting autophagy
Liraglutide prevents high glucose level induced insulinoma cells apoptosis by targeting autophagy
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DOI：
10.3760/cma.j.issn.0366-6999.20122331
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发表时间：
2013-03-05
影响因子：
6.1
通讯作者：
Xie Guang-ying
中科院分区：
文献类型：
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作者：
Chen Ze-fang;Li Yan-bo;Xie Guang-ying
背景 2 型糖尿病的病理生理学是进行性胰腺 β 细胞衰竭，从而导致胰岛素分泌减少。糖毒性通过诱导细胞凋亡导致 2 型糖尿病中 β 细胞质量减少。自噬对于维持正常胰岛结构至关重要，并通过加速β细胞中的胰岛素降解速率在维持细胞内胰岛素含量方面发挥着至关重要的作用。近年来自噬在2型糖尿病中的作用越来越受到关注。自噬控制胰腺β细胞的调控途径目前尚不清楚。我们研究的目的是评估利拉鲁肽是否可以在体外抑制胰岛素瘤细胞（INS-1细胞）的凋亡并调节自噬。方法将INS-1细胞在存在或不存在高水平葡萄糖、利拉鲁肽（长效人胰高血糖素样肽-1类似物）或3-亚腺嘌呤（3-MA）的情况下孵育24小时。使用 Cell Counting Kit-8 (CCK8) 活力测定法测量细胞活力。通过单丹磺酰尸胺 (MDC) 染色（一种用于标记自噬泡的自噬荧光化合物）和微管相关蛋白 I 轻链 3 (LC3)（自噬启动的生化标记物）的蛋白质印迹来测试 INS-1 细胞的自噬。 结果 高水平葡萄糖处理后 INS-1 细胞的活力降低。当自噬受到抑制时，INS-1 细胞的活力降低，细胞凋亡增加。与单独高糖处理的细胞相比，添加利拉鲁肽补充高糖水平的INS-1细胞活力显着增加。结论高糖处理时大鼠INS-1细胞凋亡和自噬增加，抑制自噬显着降低INS-1细胞的活力。利拉鲁肽保护 INS-1 细胞免受高葡萄糖水平诱导的细胞凋亡，并伴有自噬的显着增加，表明利拉鲁肽通过靶向自噬在 β 细胞凋亡中发挥作用。因此，自噬可能成为预防或治疗糖尿病的新靶点。
Background The pathophysiology of type 2 diabetes is progressive pancreatic beta cell failure with consequential reduced insulin secretion. Glucotoxicity results in the reduction of beta cell mass in type 2 diabetes by inducing apoptosis. Autophagy is essential for the maintenance of normal islet architecture and plays a crucial role in maintaining the intracellular insulin content by accelerating the insulin degradation rate in beta cells. Recently more attention has been paid to the effect of autophagy in type 2 diabetes. The regulatory pathway of autophagy in controlling pancreatic beta cells is still not clear. The aim of our study was to evaluate whether liraglutide can inhibit apoptosis and modulate autophagy in vitro in insulinoma cells (INS-1 cells).Methods INS-1 cells were incubated for 24 hours in the presence or absence of high levels of glucose, liraglutide (a long-acting human glucagon-like peptide-1 analogue), or 3-methyadenine (3-MA). Cell viability was measured using the Cell Counting Kit-8 (CCK8) viability assay. Autophagy of INS-1 cells was tested by monodansylcadaverine (MDC) staining, an autophagy fluorescent compound used for the labeling of autophagic vacuoles, and by Western blotting of microtubule-associated protein I light chain 3 (LC3), a biochemical markers of autophagic initiation.Results The viability of INS-1 cells was reduced after treatment with high levels of glucose. The viability of INS-1 cells was reduced and apoptosis was increased when autophagy was inhibited. The viability of INS-1 cells was significantly increased by adding liraglutide to supplement high glucose level medium compared with the cells treated with high glucose levels alone.Conclusions Apoptosis and autophagy were increased in rat INS-1 cells when treated with high level of glucose, and the viability of INS-1 cells was significantly reduced by inhibiting autophagy. Liraglutide protected INS-1 cells from high glucose level-induced apoptosis that is accompanied by a significant increase of autophagy, suggesting that liraglutide plays a role in beta cell apoptosis by targeting autophagy. Thus, autophagy may be a new target for the prevention or treatment of diabetes.