Department of Biochemical Sciences, University of Florence, Viale Morgagni 50, 50134, Florence, Italy.
J Mol Med (Berl). 2010 Jun;88(6):597-608. doi: 10.1007/s00109-010-0603-8. Epub 2010 Mar 10.
Increasing evidence indicates that cell surfaces are early interaction sites for Abeta-derived diffusible ligands (ADDLs) and neurons in Alzheimer's disease (AD) pathogenesis. Our previous data showed significant oxidative damage at the plasma membrane in fibroblasts from familial AD patients with enhanced Abeta production. Here, we report that lipid rafts, ordered membrane microdomains, are chronic mediators of Abeta-induced lipid peroxidation in SH-SY5Y human neuroblastoma cells overexpressing amyloid precursor protein (APPwt) and APPV717G genes and in fibroblasts bearing the APPV717I gene mutation. Confocal microscope analysis showed that Abeta-oxidised rafts recruit more ADDLs than corresponding domains in control cells, triggering a further increase in membrane lipid peroxidation and loss of membrane integrity. Moreover, amyloid pickup at the oxidative-damaged domains was prevented by enhanced cholesterol levels, anti-ganglioside (GM1) antibodies, the B subunit of cholera toxin and lipid raft structure alteration. An enhanced structural rigidity of the detergent-resistant domains, isolated from APPwt and APPV717G cells and exposed to ADDLs, indicates a specific perturbation of raft physicochemical features in cells facing increased amyloid assembly at the membrane surface. These data identify lipid rafts as key mediators of oxidative damage as a result of their ability to recruit aggregates to the cell surface.
越来越多的证据表明,细胞表面是阿尔茨海默病(AD)发病机制中 Abeta 衍生可扩散配体(ADDL)和神经元的早期相互作用位点。我们之前的数据表明,在具有增强的 Abeta 产生的家族性 AD 患者的成纤维细胞的质膜上存在显著的氧化损伤。在这里,我们报告脂质筏,即有序的膜微区,是 SH-SY5Y 人神经母细胞瘤细胞中 Abeta 诱导的脂质过氧化的慢性介质,这些细胞过度表达淀粉样前体蛋白(APPwt)和 APPV717G 基因,以及携带 APPV717I 基因突变的成纤维细胞。共焦显微镜分析表明,氧化的筏募集的 ADDLs 比对照细胞中相应的域多,从而引发膜脂质过氧化和膜完整性丧失的进一步增加。此外,通过增强胆固醇水平、抗神经节苷脂(GM1)抗体、霍乱毒素 B 亚基和脂质筏结构改变,可防止氧化损伤域的淀粉样蛋白摄取。从 APPwt 和 APPV717G 细胞中分离出来并暴露于 ADDLs 的去污剂抗性域的增强结构刚性表明,由于其能够将聚集物募集到质膜表面,因此质筏物理化学特性发生了特定的扰动。这些数据将脂质筏确定为氧化损伤的关键介质,因为它们能够将聚集物募集到细胞表面。