Metal oxide nanoparticles (NPs), such as ZnO, ZnFe₂O₄, and Fe₂O₃, are widely used in industry. However, little is known about the cellular pathways involved in their potential toxicity. Here, we particularly investigated the key molecular pathways that are switched on after exposure to sub-toxic doses of ZnO, ZnFe₂O₄, and Fe₂O₃ in the in vitro rat alveolar macrophages (NR8383). As in our model, the calculated IC₅₀ were respectively 16, 68, and more than 200 μg/mL for ZnO, ZnFe₂O₄, and Fe₂O₃; global gene and protein expression profiles were only analyzed after exposure to ZnO and ZnFe₂O₄ NPs. Using a rat genome microarray technology, we found that 985 and 1209 genes were significantly differentially expressed in NR8383 upon 4 h exposure to ¼ IC₅₀ of ZnO and ZnFe₂O₄ NPs, respectively. It is noteworthy that metallothioneins were overexpressed genes following exposure to both NPs. Moreover, Ingenuity Pathway Analysis revealed that the top canonical pathway disturbed in NR8383 exposed to ZnO and ZnFe₂O₄ NPs was eIF2 signaling involved in protein homeostasis. Quantitative mass spectrometry approach performed from both NR8383 cell extracts and culture supernatant indicated that 348 and 795 proteins were differentially expressed upon 24 h exposure to ¼ IC₅₀ of ZnO and ZnFe₂O₄ NPs, respectively. Bioinformatics analysis revealed that the top canonical pathways disturbed in NR8383 were involved in protein homeostasis and cholesterol biosynthesis for both exposure conditions. While VEGF signaling was specific to ZnO exposure, iron homeostasis signaling pathway was specific to ZnFe₂O₄ NPs. Overall, the study provides resource of transcriptional and proteomic markers of response to ZnO and ZnFe₂O₄ NP-induced toxicity through combined transcriptomics, proteomics, and bioinformatics approaches.

中文翻译：

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暴露于金属氧化物纳米粒子后，大鼠巨噬细胞中的蛋白质和脂质体内稳态发生改变。

诸如ZnO，ZnFe ₂ O ₄和Fe ₂ O _3的金属氧化物纳米颗粒（NPs）在工业中被广泛使用。然而，关于其潜在毒性涉及的细胞途径知之甚少。在这里，我们特别研究了在体外大鼠肺泡巨噬细胞（NR8383）中暴露于亚毒性剂量的ZnO，ZnFe ₂ O ₄和Fe ₂ O ₃后打开的关键分子途径。与我们的模型一样，对于ZnO，ZnFe ₂ O ₄和Fe ₂ O ₃，计算出的IC ₅₀分别为16、68和200μg/ mL以上。; 仅在暴露于ZnO和ZnFe ₂ O ₄ NP后才分析全局基因和蛋白质表达谱。使用大鼠基因组微阵列技术，我们发现985和1209基因在NR8383中分别暴露于ZnO和ZnFe ₂ O ₄ NP的¼IC ₅₀分别4 h时显着差异表达。值得注意的是，金属硫蛋白在暴露于两个NP后都是过表达的基因。此外，“机能路径分析”显示，在暴露于ZnO和ZnFe ₂ O _4的NR8383中，顶部经典路径受到干扰NPs是eIF2信号通路，参与蛋白质稳态。从NR8383细胞提取物和培养上清液中进行的定量质谱分析方法表明，分别在ZnO和ZnFe ₂ O ₄ NP的1/4 IC ₅₀暴露24 h后，348和795蛋白差异表达。生物信息学分析表明，在两种暴露条件下，NR8383中受干扰的最常见的经典途径均与蛋白质稳态和胆固醇生物合成有关。尽管VEGF信号转导特定于ZnO暴露，但铁稳态信号转导途径特定于ZnFe ₂ O ₄ NP。总体而言，该研究提供了对ZnO和ZnFe ₂响应的转录和蛋白质组学标记物的资源O ₄ NP通过转录组学，蛋白质组学和生物信息学方法联合诱导的毒性。