Silver nanomaterials (AgNMs) are broadly used and among the most studied nanomaterials. The underlying molecular mechanisms (e.g. protein and metabolite response) that precede phenotypical effects have been assessed to a much lesser extent. In this paper, we assess differentially expressed proteins (DEPs) and metabolites (DEMs) by high-throughput (HTP) techniques (HPLC-MS/MS with tandem mass tags, reversed-phase (RP) and hydrophilic interaction liquid chromatography (HILIC) with mass spectrometric detection). In a time series (0, 7, 14 days), the standard soil model Enchytraeus crypticus was exposed to AgNM300K and AgNO₃ at the reproduction EC20 and EC50. The impact on proteins/metabolites was clearly larger after 14 days. NM300K caused more upregulated DEPs/DEMs, more so at the EC20, whereas AgNO₃ caused a dose response increase of DEPs/DEMs. Similar pathways were activated, although often via opposite regulation (up vs down) of DEPs, hence, dissimilar mechanisms underlie the apical observed impact. Affected pathways included e.g. energy and lipid metabolism and oxidative stress. Uniquely affected by AgNO₃ was catalase, malate dehydrogenase and ATP-citrate synthase, and heat shock proteins (HSP70) and ferritin were affected by AgNM300K. The gene expression-based data in Adverse Outcome Pathway was confirmed and additional key events added, e.g. regulation of catalase and heat shock proteins were confirmed to be included. Finally, we observed (as we have seen before) that lower concentration of the NM caused higher biological impact. Data was deposited to ProteomeXchange, identifier PXD024444.

银纳米材料（AgNMs）应用广泛，是研究最多的纳米材料之一。在表型效应之前的潜在分子机制（例如蛋白质和代谢物反应）的评估程度要小得多。在本文中，我们通过高通量 (HTP) 技术（带串联质量标签的 HPLC-MS/MS、反相 (RP) 和亲水相互作用液相色谱 (HILIC)）评估差异表达的蛋白质 (DEP) 和代谢物 (DEM)质谱检测）。在时间序列（0、7、14天）中，标准土壤模型Enchytraeus crypticus暴露于 AgNM300K 和 AgNO ₃在繁殖 EC20 和 EC50。14 天后对蛋白质/代谢物的影响明显更大。NM300K 引起更多上调的 DEPs/DEMs，在 EC20 上更是如此，而 AgNO ₃引起DEPs/DEMs的剂量反应增加。类似的通路被激活，尽管通常是通过DEP 的相反调节（向上与向下），因此，不同的机制构成了观察到的顶端影响的基础。受影响的途径包括例如能量和脂质代谢以及氧化应激。独特地受 AgNO ₃影响是过氧化氢酶、苹果酸脱氢酶和 ATP-柠檬酸合酶，热休克蛋白 (HSP70) 和铁蛋白受 AgNM300K 影响。Adverse Outcome Pathway 中基于基因表达的数据得到确认，并确认添加了额外的关键事件，例如过氧化氢酶和热休克蛋白的调节。最后，我们观察到（如我们之前所见）较低浓度的 NM 会导致较高的生物影响。数据存入 ProteomeXchange，标识符为 PXD024444。