Department of Medical Physics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; Nanomedicine and Nanobiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Department of Nanomedicine, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.
Int J Biol Macromol. 2018 Jun;112:703-711. doi: 10.1016/j.ijbiomac.2018.02.025. Epub 2018 Feb 6.
Fibrillation inhibition effects of chemically and biogenically gold nanoparticles (GNPs) were investigated in vitro using human insulin as a model for fibrillation of protein. This inspection was followed using the Congo red assay, thioflavin T fluorescence measurements, transmission electron microscopy, and evaluation of cytotoxicity effects on rat pheochromocytoma PC12 cells. Biogenic GNPs were synthesized using oil extracts of Citrus aurantium L. blossoms and Rose damascena blossoms as reducing and concomitant agents. Congo red assay showed development of fibril formation of insulin at acidic media at 60°C over a period of 48h. In these circumstances, transmission electron micrographs confirmed the progress of fibril state from globular chains to amyloid. However, the results of ThT fluorescence measurements indicated a concentration-dependent inhibiting effect of chemically synthesized GNPs on insulin fibrillation in vitro, simultaneously by conversion of the formed fibrils into amorphous aggregates. Furthermore, biogenic GNPs were found to more effectively inhibit the fibril formation, compared to chemically synthesized GNPs. Accordingly, just 0.05nmolL of the biogenic GNPs showed similar inhibition property of chemically synthesized GNPs with a concentration of 10nmolL. Both types of GNPs diminished toxicity of insulin fibrils in rat pheochromocytoma PC12 cells viability.
采用刚果红法、硫代黄素 T 荧光法、透射电镜观察和对大鼠嗜铬细胞瘤 PC12 细胞的细胞毒性评价等方法,研究了化学法和生物法制备的金纳米粒子(GNPs)对人胰岛素纤维形成的抑制作用。实验以胰岛素为模型蛋白,考察了 GNPs 对蛋白纤维形成的抑制作用。生物法 GNPs 是用柑橘属橘花和大马士革玫瑰的油提取物作为还原剂和共还原剂合成的。刚果红法显示,胰岛素在 60°C 的酸性介质中,经过 48h 的时间形成纤维。在这些条件下,透射电镜证实了纤维状态从球状链到淀粉样的进展。然而,ThT 荧光法的结果表明,化学合成的 GNPs 对体外胰岛素纤维形成具有浓度依赖性的抑制作用,同时将形成的纤维转化为无定形聚集体。此外,生物法 GNPs 对纤维形成的抑制作用比化学合成的 GNPs 更有效。因此,生物法 GNPs 只需 0.05nmol/L 即可显示出与浓度为 10nmol/L 的化学合成 GNPs 相似的抑制作用。两种类型的 GNPs 均降低了胰岛素纤维在大鼠嗜铬细胞瘤 PC12 细胞活力中的毒性。