Kasi Phanindra B, Kotorman Marta, Borics Attila, Hervay Beata G, Molnar Kinga, Laszlo Lajos
Department of Biochemistry and Molecular Biology, Faculty of Science and Informatics, University of Szeged, Kozepfasor 52, H-6726 Szeged, Hungary.
Laboratory of Chemical Biology, Biological Research Centre of the Hungarian Academy of Sciences, Temesvari krt. 62, H-6726 Szeged, Hungary.
Protein Pept Lett. 2018;25(3):253-259. doi: 10.2174/0929866525666171229231226.
In case of several chronic diseases, prevention is could be more effective than treatment. Functional foods that contain significant amounts of bioactive components gained considerable attention not only in traditional but in modern medicine as well. We have investigated how P. ginseng extract inhibits the in vitro formation of amyloid-like fibrils of phenylmethylsulfonyl- trypsin (PMS-trypsin) in 60% ethanol at pH 7.0. The model system used is non-physiological, but it is capable of detecting the anti-amyloidogenic effect of the various agents.
The main objective of this study was to examine the possible inhibitory effect of ginseng extract on amyloid-like fibril formation of trypsin in aqueous ethanol.
The amyloid formation and aggregation kinetics of PMS-trypsin was studied by turbidity measurements, Congo Red (CR) binding assays, size exclusion chromatography and Electronic Circular Dichroism (ECD) measurements and the shapes of amyloid fibrils became visible by Transmission Electron Microscopy (TEM).
In the presence of 500-fold diluted P. ginseng extract in the incubation mixture, the absorption at 350 nm decreased to 47.1% after incubation for 24 h, compared relative to the sample which contained no additives. CR binding experiments suggested that the aggregates in our samples have amyloid-like properties, and P. ginseng extract inhibits the amyloid-like fibril formation of PMS-trypsin depending on concentration. Our results show that the ginseng extract does not bind to the fibrils. In the absence of P. ginseng extracts large sized colloid aggregates were abundant. Adding P. ginseng extracts to our samples decreased the light dispersion of the solution. This is due to the decrease of the rate of the aggregation or to the smaller size of the aggregates evolved. Our results show that the presence of ginseng extract helps to maintain the native structure of the protein. In the presence of 500-fold diluted P. ginseng extract, TEM images demonstrated, that P. ginseng extract has inhibitory effect on the formation of amyloid-like fibrils of PMS-trypsin.
The results indicated that P. ginseng extract significantly inhibits the formation of amyloid-like fibrils of PMS-trypsin in aqueous ethanol, and helps to maintain the native structure of the protein. The rate of inhibition depends on concentration. P. ginseng extract is an efficient antiamyloidogenic agent.
对于几种慢性疾病，预防可能比治疗更有效。含有大量生物活性成分的功能性食品不仅在传统医学中，而且在现代医学中也受到了相当多的关注。我们研究了人参提取物如何在pH 7.0的60%乙醇中抑制苯甲基磺酰 - 胰蛋白酶（PMS - 胰蛋白酶）淀粉样纤维的体外形成。所使用的模型系统是非生理性的，但它能够检测各种试剂的抗淀粉样蛋白生成作用。
本研究的主要目的是检测人参提取物对胰蛋白酶在乙醇水溶液中淀粉样纤维形成的可能抑制作用。
通过浊度测量、刚果红（CR）结合试验、尺寸排阻色谱和电子圆二色性（ECD）测量研究PMS - 胰蛋白酶的淀粉样蛋白形成和聚集动力学，并通过透射电子显微镜（TEM）观察淀粉样纤维的形状。
在孵育混合物中存在500倍稀释的人参提取物时，孵育24小时后，相对于不含添加剂的样品，350nm处的吸光度降至47.1%。CR结合实验表明我们样品中的聚集体具有淀粉样蛋白样特性，人参提取物根据浓度抑制PMS - 胰蛋白酶的淀粉样纤维形成。我们的结果表明人参提取物不与纤维结合。在没有人参与提取物的情况下，大量存在大尺寸的胶体聚集体。向我们的样品中添加人参提取物降低了溶液的光散射。这是由于聚集速率降低或所形成聚集体的尺寸较小。我们的结果表明人参提取物的存在有助于维持蛋白质的天然结构。在存在500倍稀释的人参提取物的情况下，TEM图像表明，人参提取物对PMS - 胰蛋白酶淀粉样纤维的形成具有抑制作用。
结果表明人参提取物在乙醇水溶液中显著抑制PMS - 胰蛋白酶淀粉样纤维的形成，并有助于维持蛋白质的天然结构。抑制率取决于浓度。人参提取物是一种有效的抗淀粉样蛋白生成剂。
