Department of Geriatrics, The Second Hospital of Hebei Medical University, 050000 Shijiazhuang, Hebei, China.
J Integr Neurosci. 2023 Feb 14;22(2):33. doi: 10.31083/j.jin2202033.
Ginkgolide B (GB) possesses anti-inflammatory, antioxidant, and anti-apoptotic properties against neurotoxicity induced by amyloid beta (Aβ), but the potential neuroprotective effects of GB in Alzheimer's therapies remain elusive. We aimed to conduct proteomic analysis of Aβ1-42 induced cell injury with GB pretreatment to uncover the underlying pharmacological mechanisms of GB.
Tandem mass tag (TMT) labeled liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was applied to analyze protein expression in Aβ1-42 induced mouse neuroblastoma N2a cells with or without GB pretreatment. Proteins with fold change >1.5 and < 0.1 from two independent experiments were regarded as differentially expressed proteins (DEPs). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to analyze the functional annotation information of DEPs. Two key proteins osteopontin (SPP1) and ferritin heavy chain 1 (FTH1) were validated in another three samples using western blot and quantitative real-time PCR.
We identified a total of 61 DEPs in GB treated N2a cells, including 42 upregulated and 19 downregulated proteins. Bioinformatic analysis showed that DEPs mainly participated in the regulation of cell death and ferroptosis by down-regulating SPP1 protein and up-regulating FTH1 protein.
Our findings demonstrate that GB treatment provides neuroprotective effects on Aβ1-42 induced cell injury, which may be related to the regulation of cell death and ferroptosis. The research puts forward new insights into the potential protein targets of GB in the treatment of Alzheimer's disease (AD).
银杏内酯 B（GB）具有抗炎、抗氧化和抗细胞凋亡作用，可对抗淀粉样蛋白β（Aβ）诱导的神经毒性，但 GB 在阿尔茨海默病治疗中的潜在神经保护作用仍不清楚。我们旨在进行蛋白质组学分析，研究 GB 预处理对 Aβ1-42 诱导的细胞损伤的影响，以揭示 GB 的潜在药理机制。
采用串联质量标签（TMT）标记液相色谱-串联质谱（LC-MS/MS）法分析 Aβ1-42 诱导的小鼠神经母细胞瘤 N2a 细胞中有无 GB 预处理的蛋白表达。两个独立实验中 fold change >1.5 且 < 0.1 的蛋白被视为差异表达蛋白（DEPs）。对 DEPs 进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析，以分析 DEPs 的功能注释信息。使用 Western blot 和定量实时 PCR 在另外三个样本中验证了两个关键蛋白骨桥蛋白（SPP1）和铁蛋白重链 1（FTH1）。
我们在 GB 处理的 N2a 细胞中总共鉴定出 61 个 DEPs，包括 42 个上调蛋白和 19 个下调蛋白。生物信息学分析表明，DEPs 主要通过下调 SPP1 蛋白和上调 FTH1 蛋白来参与细胞死亡和铁死亡的调节。
我们的研究结果表明，GB 治疗对 Aβ1-42 诱导的细胞损伤具有神经保护作用，这可能与细胞死亡和铁死亡的调节有关。该研究为 GB 治疗阿尔茨海默病（AD）的潜在蛋白靶标提供了新的见解。
