Wang Zhifang, Zhou Jingpei, Zhang Bin, Xu Zhanqiong, Wang Haoyu, Sun Quan, Wang Nanbu
Guangzhou University of Chinese Medicine, Guangzhou, China.
Guangzhou University of Traditional Chinese Medicine First Affiliated Hospital, Guangzhou, China.
Behav Brain Res. 2024 Apr 12;463:114896. doi: 10.1016/j.bbr.2024.114896. Epub 2024 Feb 4.
The primary aim of this study was to examine the correlation between the formation of Aβ plaques and autophagy, which is regulated by β-asarone and the lncRNA BACE1-AS. Additionally, the study sought to explore potential targets of the drug in inhibiting the deposition of toxic AD-related proteins and restoring impaired mitochondrial and autophagic functions. SHY5Y cells were utilized to construct a stable Alzheimer's disease (AD) model, followed by the utilization of interference and overexpression lentiviruses targeting BACE1-AS to establish a cell model. The cells were categorized into five groups, including a normal group, siRNA/BACE1 group, and β-asarone group. The fluorescence quantitative PCR technique was employed to assess the disparity in BACE1 mRNA expression, while changes in immunofluorescence (IF) were observed to determine the stable interference titre and action time of the lentiviruses. Additionally, western blotting (WB) and fluorescence quantitative PCR were employed to evaluate the expression of proteins and mRNAs associated with AD and autophagy. The findings demonstrated a significant elevation in BACE1 expression levels in brain tissue among individuals with AD compared to those without the condition. Moreover, the results indicated that the introduction of β-asarone led to an increase in the expression of the BACE1-AS gene in the cell group transfected with plasmid H12732. Furthermore, it was observed that β-asarone enhanced the expression levels of shRNA and BACE1 after 72 h. In contrast, β-asarone suppressed the expression of PS1, Aβ, BACE1, APP, and p62, while promoting the expression of syn, LC3 I/II, and Beclin-1. Based on these findings, it can be concluded that β-Asarone exerts a comprehensive influence on the expression of proteins associated with AD and synaptic function. β-Asarone exhibits the potential to mitigate Aβ deposition by impeding the expression of lncBACE1, thereby facilitating autophagy through the suppression of BACE1's inhibitory impact on autophagy. This complements the self-enhancing effect of autophagy.
本研究的主要目的是检测β-细辛醚和长链非编码RNA BACE1-AS调控的淀粉样β蛋白（Aβ）斑块形成与自噬之间的相关性。此外，该研究还试图探索该药物在抑制有毒的阿尔茨海默病（AD）相关蛋白沉积以及恢复受损的线粒体和自噬功能方面的潜在靶点。利用SHY5Y细胞构建稳定的AD模型，随后利用靶向BACE1-AS的干扰和过表达慢病毒建立细胞模型。细胞分为五组，包括正常组、siRNA/BACE1组和β-细辛醚组。采用荧光定量PCR技术评估BACE1 mRNA表达差异，同时观察免疫荧光（IF）变化以确定慢病毒的稳定干扰效价和作用时间。此外，采用蛋白质免疫印迹法（WB）和荧光定量PCR评估与AD和自噬相关的蛋白质和mRNA表达。结果表明，与非AD患者相比，AD患者脑组织中BACE1表达水平显著升高。此外，结果表明，在转染质粒H12732的细胞组中，β-细辛醚导致BACE1-AS基因表达增加。此外，观察到β-细辛醚在72小时后增强了shRNA和BACE1的表达水平。相反，β-细辛醚抑制了早老素1（PS1）、Aβ、BACE1、淀粉样前体蛋白（APP）和p62的表达，同时促进了突触素（syn）、微管相关蛋白轻链3（LC3 I/II）和Beclin-1的表达。基于这些发现，可以得出结论，β-细辛醚对与AD和突触功能相关的蛋白质表达具有全面影响。β-细辛醚具有通过阻碍lncBACE1的表达来减轻Aβ沉积的潜力，从而通过抑制BACE1对自噬的抑制作用促进自噬。这补充了自噬的自我增强作用。
