Department of Neurology, The First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, 550002 Guiyang, Guizhou, China.
The First School of Clinical Medicine of Guizhou University of Traditional Chinese Medicine, 550001 Guiyang, Guizhou, China.
Actas Esp Psiquiatr. 2024 Apr;52(2):83-98. doi: 10.62641/aep.v52i2.1601.
Vascular dementia (VaD) is a prevalent neurodegenerative disease characterized by cognitive impairment due to cerebrovascular factors, affecting a significant portion of the aging population and highlighting the critical need to understand specific targets and mechanisms for effective prevention and treatment strategies. We aimed to identify pathways and crucial genes involved in the progression of VaD through bioinformatics analysis and subsequently validate these findings.
We conducted differential expression analysis, Weighted Gene Co-expression Network Analysis (WGCNA), Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and Protein-Protein Interaction (PPI) analysis. We utilized pheochromocytoma 12 (PC12) cells to create an in vitro oxygen-glucose deprivation (OGD) model. We investigated the impact of overexpression and interference of adrenoceptor alpha 1D (ADRA1D) on OGD PC12 cells using TdT-mediated dUTP nick-end labeling (TUNEL), reverse transcription-quantitative polymerase chain reaction (RT-qPCR), western blot (WB), and Fluo-3-pentaacetoxymethyl ester (Fluo-3 AM) analysis.
We found 187 differentially expressed genes (DEGs) in the red module that were strongly associated with VaD and were primarily enriched in vasoconstriction, G protein-coupled amine receptor activity, and neuroactive ligand-receptor interaction, mitogen-activated protein kinase (MAPK) signaling pathway, and cell adhesion. Among these pathways, we identified ADRA1D as a gene shared by vasoconstriction, G protein-coupled amine receptor activity, and neuroactive ligand-receptor interaction. The TUNEL assay revealed a significant decrease in PC12 cell apoptosis with ADRA1D overexpression (p < 0.01) and a significant increase in apoptosis upon silencing ADRA1D (p < 0.01). RT-qPCR and WB analysis revealed elevated ADRA1D expression (p < 0.001) and decreased phospholipase C beta (PLCβ) and inositol 1,4,5-trisphosphate receptor (IP3R) expression (p < 0.05) with ADRA1D overexpression. Moreover, the Fluo-3 AM assessment indicated significantly lower intracellular Ca2+ levels with ADRA1D overexpression (p < 0.001). Conversely, interference with ADRA1D yielded opposite results.
Our study provides a new perspective on the pathogenic mechanisms of VaD and potential avenues for therapeutic intervention. The results highlight the role of ADRA1D in modulating cellular responses to OGD and VaD, suggesting its potential as a target for VaD treatment.
血管性痴呆（VaD）是一种常见的神经退行性疾病，其特征是由于脑血管因素导致的认知障碍，影响着相当一部分老年人群体，凸显了深入了解特定靶点和机制以制定有效预防和治疗策略的重要性。我们旨在通过生物信息学分析来鉴定 VaD 进展过程中的相关通路和关键基因，并随后对这些发现进行验证。
我们进行了差异表达分析、加权基因共表达网络分析（WGCNA）、基因本体论（GO）、京都基因与基因组百科全书（KEGG）通路富集分析和蛋白质-蛋白质相互作用（PPI）分析。我们利用嗜铬细胞瘤 12 细胞（PC12）建立了体外氧葡萄糖剥夺（OGD）模型。我们通过末端转移酶介导的 dUTP 缺口末端标记（TUNEL）、逆转录定量聚合酶链反应（RT-qPCR）、蛋白质印迹（WB）和 Fluo-3-五乙酸甲酯（Fluo-3 AM）分析，研究了过表达和干扰肾上腺素能受体 alpha 1D（ADRA1D）对 OGD PC12 细胞的影响。
我们在与 VaD 密切相关的红色模块中发现了 187 个差异表达基因（DEGs），这些基因主要富集在血管收缩、G 蛋白偶联胺受体活性、神经活性配体-受体相互作用、丝裂原活化蛋白激酶（MAPK）信号通路和细胞黏附。在这些通路中，我们确定 ADRA1D 是血管收缩、G 蛋白偶联胺受体活性和神经活性配体-受体相互作用的共享基因。TUNEL 检测结果显示，ADRA1D 过表达时 PC12 细胞凋亡显著减少（p < 0.01），ADRA1D 沉默时凋亡显著增加（p < 0.01）。RT-qPCR 和 WB 分析显示 ADRA1D 过表达时 ADRA1D 表达升高（p < 0.001），磷脂酶 Cβ（PLCβ）和三磷酸肌醇受体（IP3R）表达降低（p < 0.05）。此外，Fluo-3 AM 评估显示 ADRA1D 过表达时细胞内 Ca2+水平显著降低（p < 0.001）。相反，干扰 ADRA1D 则产生相反的结果。
本研究为 VaD 的发病机制提供了新的视角，并为治疗干预提供了潜在的途径。结果强调了 ADRA1D 在调节细胞对 OGD 和 VaD 的反应中的作用，提示其可能成为 VaD 治疗的靶点。
