Effect of Danggui Shaoyaosan on Copper Ion-mediated Aβ Aggregation in AD Cell Model / 中国实验方剂学杂志

ABSTRACT

Objective:To investigate the effect of copper ion(Cu2+) on the aggregation and neurotoxicity of Aβ, and affirm the role of Danggui Shaoyaosan in vitro,the Neuroblastoma (SH-SY5Y) cells treated with β-amyloid 1-42 (Aβ1-42) and Cu2+ were used as a vitro models of Alzheimer's disease(AD). Method:Aβ 1-42 (20 μmol·L-1) was reacted with different concentrations of copper sulfate (CuSO4,20,40 μmol·L-1), and then the thioflavine T (ThT) staining method was used to detect the Aβ aggregation state. The Aβ aggregation status was also detected by ThT staining in the Aβ1-42-Cu2+ group(20+20 μmol·L-1), and Danggui Shaoyaosan groups(1.6,3.2,6.4 mg·L-1).The SH-SY5Y cells were cultured and incubated with different concentrations of Aβ1-42(1.25,2.5,5,10,20,40 μmol·L-1) and Danggui Shaoyaosan(1.6, 3.2, 6.4,12.8 mg·L-1) for 24 h. Subsequently, SH-SY5Y cells were incubated with Aβ1-42 (20 μmol·L-1) and CuSO4(20 μmol·L-1) in the Aβ1-42-Cu2+ group, and incubated with Aβ1-42 (20 μmol·L-1), Danggui Shaoyaosan (1.6 mg·L-1) and CuSO4 (20 μmol·L-1) in Danggui Shaoyaosan group. Control group was added with the medium. After 24 h of co-action, the cell viability was detected by the methylthiazolyl tetrazolium (MTT) assay. The morphology of the cells was photographed by microscopy. The intracellular extracellular Aβ1-42 aggregation was detected by Western blot. Result:Cu2+ and Aβ1-42 bound to more and larger Aβ aggregates compared with the Aβ1-42 group. Compared with the normal group, cell viability was significantly reduced (PPβ1-42 aggregation was increased(PPβ-Cu2+ (PPβ1-42 protein (PPConclusion:Cu2+ can increase the aggregation and toxicity of Aβ; Danggui Shaoyaosan can significantly reduce the damage of SH-SY5Y cells induced by Cu2+-mediated Aβ aggregation, promote Aβ endocytosis, reduce extracellular Aβ aggregation and increase cell viability.