Institute of Biochemistry and Biophysics, PO Box 13145-1384, University of Tehran, Enghelab Ave, Tehran, Iran.
Eur J Pharmacol. 2013 May 5;707(1-3):17-25. doi: 10.1016/j.ejphar.2013.03.017. Epub 2013 Mar 23.
Aggregation of human islet amyloid polypeptide (IAPP) into pancreatic fibrillar deposits has been postulated to be one of the main contributors to impaired insulin secretion and pancreatic β-cell death in approximately 90% of type 2 diabetic patients. So, compounds that prevent cytotoxic protein/polypeptide self-assembly and amyloidogenesis are considered as novel therapeutic agents against this disease. In this study, using thioflavin-T (ThT) and Anilinonaphthalene-8-sulfonic acid (ANS) fluorescence assays, transmission electron microscopy (TEM) and docking studies, we investigated whether EUK-8 and EUK-134, two salen derivatives with proven antioxidants activities, could interfere with the conversion of synthetic human amylin to its insoluble amyloid form. Spectroscopy and electron microscopy data indicated that incubation of human amylin with either EUK-8 or EUK-134 significantly inhibited amyloid formation at two molar ratios of 1:1 and 5:1 (drugs to protein). In addition, [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assay showed that treatment of SK-N-MC cells with the pre-formed fibrils in the presence of compounds at drug-to-protein molar ratios of 1:1 and 5:1, dramatically increased the viability of cells compared to the only fibrils formed-treated SK-N-MC cells. Docking results also demonstrated that the aromatic rings of EUK-8 and EUK-134 interact with the hydrophobic region (23-25) of IAPP via Van der Waals interactions. Based on these results and the proven antioxidant properties of these compounds, it could be concluded that these compounds might provide a novel approach to prevent islet amyloid deposition in β-cells and provide useful information for developing other novel compounds for the treatment of type 2 diabetes.
人胰岛淀粉样多肽(IAPP)聚集形成胰腺纤维状沉积物被认为是导致大约 90%的 2 型糖尿病患者胰岛素分泌受损和胰岛β细胞死亡的主要原因之一。因此,预防细胞毒性蛋白/多肽自组装和淀粉样变性的化合物被认为是治疗这种疾病的新型治疗剂。在这项研究中,我们使用硫黄素-T(ThT)和蒽酮萘-8-磺酸(ANS)荧光分析、透射电子显微镜(TEM)和对接研究,研究了两种已被证明具有抗氧化活性的水杨醛衍生物 EUK-8 和 EUK-134 是否可以干扰合成人胰岛淀粉样肽向其不溶性淀粉样形式的转化。光谱和电子显微镜数据表明,在摩尔比为 1:1 和 5:1(药物与蛋白)的情况下,EUK-8 或 EUK-134 与人胰岛淀粉样肽孵育可显著抑制淀粉样形成。此外,[3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴化物](MTT)测定法表明,与仅形成的纤维相比,在用化合物处理 SK-N-MC 细胞存在前体纤维时,在药物与蛋白摩尔比为 1:1 和 5:1 的情况下,细胞活力显著增加。对接结果还表明,EUK-8 和 EUK-134 的芳环通过范德华相互作用与 IAPP 的疏水区(23-25)相互作用。基于这些结果和这些化合物已被证明的抗氧化特性,可以得出结论,这些化合物可能为防止β细胞中胰岛淀粉样沉积提供一种新方法,并为开发其他用于治疗 2 型糖尿病的新型化合物提供有用信息。