Institute of Traditional Chinese Medicine, Chengde Medical College, Chengde, 067000, P.R. China.
College of Integrated Traditional Chinese and Western Medicine, Hebei University of Chinese Medicine, Shijiazhuang, 050200, P.R. China.
Comb Chem High Throughput Screen. 2024;27(14):2125-2139. doi: 10.2174/0113862073285497240226061936.
An analysis of bioinformatics and cell experiments was performed to verify the relationship between gasdermin D (GSDMD), an executive protein of pyroptosis, and Alzheimer's disease (AD).
The training set GSE33000 was utilized to identify differentially expressed genes (DEGs) in both the AD group and control group, as well as in the GSDMD protein high/low expression group. Subsequently, the weighted gene co-expression network analysis (WGCNA) and the least absolute shrinkage and selection operator (LASSO) regression analysis were conducted, followed by the selection of the key genes for the subsequent Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The association between GSDMD and AD was assessed and confirmed in the training set GSE33000, as well as in the validation sets GSE5281 and GSE48350. Immunofluorescence (IF) was employed to detect the myelin basic protein (MBP), a distinctive protein found in the rat oligodendrocytes (OLN-93 cells). A range of concentrations (1-15 μmol/L) of β-amyloid 1-42 (Aβ) were exposed to the cells, and the subsequent observations were made regarding cell morphology. Additionally, the assessments were conducted to evaluate the cell viability, the lactate dehydrogenase (LDH) release, the cell membrane permeability, and the GSDMD protein expression.
A total of 7,492 DEGs were screened using GSE33000. Subsequently, WGCNA analysis identified 19 genes that exhibited the strongest correlation with clinical traits in AD. Additionally, LASSO regression analysis identified 13 key genes, including GSDMD, AFF1, and ATOH8. Furthermore, the investigation revealed that the key genes were associated with cellular inflammation based on GO and KEGG analyses. Moreover, the area under the curve (AUC) values for the key genes in the training and validation sets were determined to be 0.95 and 0.70, respectively. Significantly, GSDMD demonstrated elevated levels of expression in AD across both datasets. The positivity of MBP expression in cells exceeded 95%. As the concentration of Aβ action gradually escalated, the detrimental effects on cells progressively intensified, resulting in a gradual decline in cell survival rate, accompanied by an increase in lactate dehydrogenase release, cell membrane permeability, and GSDMD protein expression.
The association between GSDMD and AD has been observed, and it has been found that Aβ can induce a significant upregulation of GSDMD in OLN-93 cells. This suggests that Aβ has the potential to induce cellular pyroptosis and can serve as a valuable cellular pyroptosis model for the study of AD.
通过生物信息学和细胞实验分析，验证细胞焦亡执行蛋白 GSDMD 与阿尔茨海默病（AD）之间的关系。
利用 GSE33000 训练集识别 AD 组和对照组以及 GSDMD 蛋白高/低表达组之间的差异表达基因（DEGs）。然后，进行加权基因共表达网络分析（WGCNA）和最小绝对收缩和选择算子（LASSO）回归分析，选择后续进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析的关键基因。在 GSE33000 训练集以及验证集 GSE5281 和 GSE48350 中评估和验证 GSDMD 与 AD 的关联。采用免疫荧光（IF）检测髓鞘碱性蛋白（MBP），这是大鼠少突胶质细胞（OLN-93 细胞）中的一种特征蛋白。将不同浓度（1-15μmol/L）的β-淀粉样蛋白 1-42（Aβ）暴露于细胞中，观察细胞形态的变化。此外，还评估了细胞活力、乳酸脱氢酶（LDH）释放、细胞膜通透性和 GSDMD 蛋白表达。
使用 GSE33000 筛选出 7492 个 DEGs。然后，WGCNA 分析确定了与 AD 临床特征相关性最强的 19 个基因。此外，LASSO 回归分析确定了 13 个关键基因，包括 GSDMD、AFF1 和 ATOH8。进一步研究表明，基于 GO 和 KEGG 分析，这些关键基因与细胞炎症有关。此外，在训练和验证集中，关键基因的曲线下面积（AUC）值分别为 0.95 和 0.70。值得注意的是，GSDMD 在两个数据集的 AD 中均表现出高水平的表达。细胞中 MBP 表达的阳性率超过 95%。随着 Aβ 作用浓度逐渐升高，对细胞的损害作用逐渐增强，细胞存活率逐渐下降，同时乳酸脱氢酶释放、细胞膜通透性和 GSDMD 蛋白表达增加。
观察到 GSDMD 与 AD 之间的关联，并发现 Aβ 可显著上调 OLN-93 细胞中的 GSDMD。这表明 Aβ 具有诱导细胞焦亡的潜力，可作为研究 AD 的有价值的细胞焦亡模型。
