Although the toxicity and molecular mechanisms of graphene oxide (GO) have been reported for several cell types, no proteomic study of GO has yet been conducted on macrophage cells. In this study, we used proteomics based on stable isotope labeling with amino acids in cell culture (SILAC) to quantify the proteomic changes in macrophage RAW 264.7 cells following GO treatment. We found 73 proteins that were significantly dysregulated after GO treatment. The down-regulated proteins included many ribosomal subunit proteins, indicating that GO affected cell growth. The most elevated proteins were lipoprotein lipase (LPL) and lysozyme 1 (LYZ1) which have not been reported before, and both can be used as candidate markers for GO exposure. Further enrichment analysis of the up-regulated proteins indicated these proteins are associated with the integrin complex and membrane rafts, as well as with two signal pathways: the phagosome and steroid biosynthesis pathways. We confirmed a GO concentration-dependent increase in membrane rafts and the production of phagosomes. GO exposure also induced necrotic cell death and an inflammation response in RAW 264.7 cells. We also observed an increase in the oxidative stress response (ROS) and autophagy, and the results suggest that ROS induced autophagy by the ROS-NRF2-P62 pathway.

中文翻译：

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暴露于氧化石墨烯的RAW264.7巨噬细胞的蛋白质组学分析：洞悉急性细胞反应。

尽管已经报道了氧化石墨烯（GO）的毒性和分子机制对几种细胞类型的影响，但尚未对巨噬细胞进行GO的蛋白质组学研究。在这项研究中，我们使用基于细胞培养物中氨基酸的稳定同位素标记（SILAC）的蛋白质组学来量化GO处理后巨噬细胞RAW 264.7细胞的蛋白质组学变化。我们发现73种蛋白质在GO处理后显着失调。下调的蛋白包括许多核糖体亚基蛋白，表明GO影响了细胞的生长。升高最多的蛋白是脂蛋白脂肪酶（LPL）和溶菌酶1（LYZ1），以前没有报道过，它们都可以用作GO暴露的候选标记。对上调蛋白的进一步富集分析表明，这些蛋白与整联蛋白复合物和膜筏以及两个信号途径有关：吞噬体和类固醇生物合成途径。我们证实了膜筏和吞噬体的产量随浓度的增加而增加。GO暴露还会在RAW 264.7细胞中引起坏死细胞死亡和炎症反应。我们还观察到氧化应激反应（ROS）和自噬的增加，结果表明ROS通过ROS-NRF2-P62途径诱导自噬。GO暴露还会在RAW 264.7细胞中引起坏死细胞死亡和炎症反应。我们还观察到了氧化应激反应（ROS）和自噬的增加，结果表明ROS通过ROS-NRF2-P62途径诱导自噬。GO暴露还会在RAW 264.7细胞中引起坏死细胞死亡和炎症反应。我们还观察到了氧化应激反应（ROS）和自噬的增加，结果表明ROS通过ROS-NRF2-P62途径诱导自噬。