Department of Biophysics, Faculty of Medicine, Cumhuriyet University, Sivas, Turkey.
Department of Biophysics, Faculty of Medicine, Ankara University, Ankara, Turkey.
Mol Cell Biochem. 2019 Nov;461(1-2):65-72. doi: 10.1007/s11010-019-03590-z. Epub 2019 Jul 20.
Azoramide is identified as a new compound with the dual properties for the improvement of ER-folding capacity in various cells as well as for the treatment of T2DM. Although the effect of azoramide in glucose-homeostasis in mammalians is not known very well, a limited number of experimental studies showed that it could improve the insulin sensitivity in genetically obese mice. Therefore, here, we aimed to investigate the direct effect of azoramide on insulin signaling in insulin-resistant (IR) cardiomyocytes using IR-modelled ventricular cardiomyocytes. This model was established in H9c2 cells using palmitic acid incubation (50-μM for 24-h). The development of IR in cells was verified by monitoring the cellular 2-DG6P uptake assays in these treated cells. The 2-DG6P uptake was 50% less in the IR-cells compared to the control cells, while azoramide treatment (20-μM for 48-h) could prevent fully that decrease. In addition, azoramide treatment markedly preserved the IR-induced less ATP production and high-ROS production in these IR-cells. Furthermore, this treatment prevented the functional changes in mitochondria characterized by depolarized mitochondrial membrane potential and mitochondrial fusion or fusion-related protein levels as well as cellular ATP level. Moreover, this treatment provided marked protection against IR-associated changes in the insulin signaling pathway in cells, including recovery in the phosphorylation of IRS1 and Akt as well as the protein level of GLUT4 and Akt. Our present results, for the first time, demonstrated that azoramide plays an important protective role in IR-cardiomyocytes, at most, protective action on mitochondria. Therefore, one can suggest that azoramide, as a novel regulator, can provide direct cardioprotection in the IR-heart, at most, via affecting mitochondria and can be a good candidate as a new drug for the treatment of IR-associated cardiovascular disorders in mammalians with systemic IR.
阿佐酰胺被鉴定为一种具有双重特性的新型化合物，可改善各种细胞中的 ER 折叠能力，并用于治疗 T2DM。尽管阿佐酰胺在哺乳动物葡萄糖稳态中的作用尚不清楚，但少数实验研究表明，它可以改善遗传性肥胖小鼠的胰岛素敏感性。因此，在这里，我们旨在使用 IR 模型心室心肌细胞研究阿佐酰胺对胰岛素抵抗（IR）心肌细胞中胰岛素信号的直接影响。该模型是通过在 H9c2 细胞中用棕榈酸孵育（50-μM，24 小时）建立的。通过监测这些处理细胞中的细胞 2-DG6P 摄取实验来验证细胞中的 IR 发展。与对照细胞相比，IR 细胞中的 2-DG6P 摄取减少了 50％，而阿佐酰胺处理（20-μM，48 小时）可以完全阻止这种减少。此外，阿佐酰胺处理还显着防止了 IR 诱导的这些 IR 细胞中 ATP 产生减少和高 ROS 产生。此外，这种处理防止了线粒体功能变化，其特征为线粒体膜电位去极化和线粒体融合或融合相关蛋白水平以及细胞 ATP 水平降低。此外，这种处理对细胞中与 IR 相关的胰岛素信号通路的变化提供了明显的保护作用，包括 IRS1 和 Akt 的磷酸化以及 GLUT4 和 Akt 的蛋白水平的恢复。我们的研究结果首次表明，阿佐酰胺在 IR 心肌细胞中发挥重要的保护作用，最多对线粒体起保护作用。因此，可以认为阿佐酰胺作为一种新型调节剂，可通过影响线粒体直接提供 IR 心脏的心脏保护作用，并可成为治疗哺乳动物全身性 IR 相关心血管疾病的新药候选物。
