Aβ (β-amyloid) deposition and abnormal transport were suggested to be risk factors for Alzheimer’s disease (AD). Zhenxin Xingshui Yizhi Fang (XSF), an ancient prescription in traditional Chinese medicine, was first recorded in Qianjin Yifang for treating palpitation, hypnosia, amnesia. It is reported that XSF could improve mice learning memory ability, reduce the deposition of senile plaques in hippocampus of rat brain. In this study, the neuroprotective effect of XSF against Aβ-induced apoptosis in cultured human brain microvascular endothelial cells (HBMEC) and its potential mechanism were investigated.
HBMEC cells were treated with Aβ to established neurotoxic cell model. After that, the cells were treated with 125, 250, 500 μg/mL XSF to observe the protective effect. The viability of HBMEC cells were evaluated by MTT assay, the Aβ-induced apoptosis was characterized by Hoechst-33258 and the activity of cysteinyl aspartate specific proteinase-3. The expression level of Aβ in cells induced by Aβ was measured by human Aβ kit. Protein and mRNA expression levels of advanced glycation end products (RAGE), low density lipoprotein receptor-related protein 1 (LRP1), glucose transporter 1 and 3 (GLUT1 and GLUT3) were assayed by capillary electrophoresis immunoassay and quantitative real-time polymerase chain reaction analyses.
In Aβ induced neurotoxic cells, the percentage of apoptotic cells, the concentration of Aβ and CASPASE-3 activity, protein and mRNA expression levels of RAGE increased significantly, but that of LRP1, GLUT1 and GLUT3 significantly decreased. XSF could inhibit the apoptotic of cells, reduced the concentration of Aβ and CASPASE-3 expression, downregulate RAGE and upregulate LRP1, GLUT1 and GLUT3 expression.
The results suggest that XSF can reduce the cytotoxicity of HBMEC induced by Aβ, inhibit apoptosis, and regulate the transport of Aβ on BBB and energy metabolism disorder in HBMEC.
Copyright © 2020. Published by Elsevier B.V.