Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
Int J Mol Sci. 2021 Nov 26;22(23):12780. doi: 10.3390/ijms222312780.
Significant research on Alzheimer's disease (AD) has demonstrated that amyloid β (Aβ) oligomers are toxic molecules against neural cells. Thus, determining the generation mechanism of toxic Aβ oligomers is crucial for understanding AD pathogenesis. Aβ fibrils were reported to be disaggregated by treatment with small compounds, such as epigallocatechin gallate (EGCG) and dopamine (DA), and a loss of fibril shape and decrease in cytotoxicity were observed. However, the characteristics of intermediate products during the fibril disaggregation process are poorly understood. In this study, we found that cytotoxic Aβ aggregates are generated during a moderate disaggregation process of Aβ fibrils. A cytotoxicity assay revealed that Aβ fibrils incubated with a low concentration of EGCG and DA showed higher cytotoxicity than Aβ fibrils alone. Atomic force microscopy imaging and circular dichroism spectrometry showed that short and narrow protofilaments, which were highly stable in the β-sheet structure, were abundant in these moderately disaggregated samples. These results indicate that toxic Aβ protofilaments are generated during disaggregation from amyloid fibrils, suggesting that disaggregation of Aβ fibrils by small compounds may be one of the possible mechanisms for the generation of toxic Aβ aggregates in the brain.
阿尔茨海默病(AD)的大量研究表明,淀粉样β(Aβ)寡聚体是针对神经细胞的毒性分子。因此,确定有毒 Aβ 寡聚体的产生机制对于理解 AD 的发病机制至关重要。有报道称,Aβ 原纤维可以通过小化合物如表没食子儿茶素没食子酸酯(EGCG)和多巴胺(DA)的处理而解聚,并且观察到纤维形状丧失和细胞毒性降低。然而,对于纤维解聚过程中中间产物的特征了解甚少。在本研究中,我们发现 Aβ 原纤维的适度解聚过程中会产生细胞毒性 Aβ 聚集物。细胞毒性测定表明,与单独的 Aβ 原纤维相比,用低浓度 EGCG 和 DA 孵育的 Aβ 原纤维表现出更高的细胞毒性。原子力显微镜成像和圆二色性光谱表明,这些适度解聚的样品中富含高度稳定在β-折叠结构中的短而窄的原纤维。这些结果表明,在从淀粉样纤维解聚过程中产生了有毒的 Aβ 原纤维,这表明小分子对 Aβ 纤维的解聚可能是大脑中有毒 Aβ 聚集物产生的可能机制之一。