Cytotoxic and Proinflammatory Effects of Metal-Based Nanoparticles on THP‑1 Monocytes Characterized by Combined Proteomics Approaches

Thorough characterization of toxic effects of nanoparticles (NP) is desirable due to the increasing risk of potential environmental contamination by NP. In the current study, we combined three recently developed proteomics approaches to assess the effects of Au, CuO, and CdTe NP on the innate immune system. The human monocyte cell line THP-1 was employed as a model. The anticancer drugs camptothecin and doxorubicin were used as positive controls for cell death, and lipopolysaccharide was chosen as a positive control for proinflammatory activation. Despite equivalent overall toxicity effect (50 ± 10% dead cells), the three NP induced distinctly different proteomics signatures, with the strongest effect being induced by CdTe NP, followed by CuO and gold NP. The CdTe toxicity mechanism involves down-regulation of topoisomerases. The effect of CuO NP is most reminiscent of oxidative stress and involves up-regulation of proteins involved in heat response. The gold NP induced up-regulation of the inflammatory mediator, NF-κB, and its inhibitor TIPE2 was identified as a direct target of gold NP. Furthermore, gold NP triggered activation of NF-κB as evidenced by phosphorylation of the p65 subunit. Overall, the combined proteomics approach described here can be used to characterize the effects of NP on immune cells.

鉴于纳米颗粒（nanoparticles, NP）潜在环境污染风险日益升高，对其毒性效应开展全面表征具有重要意义。本研究结合三种新近开发的蛋白质组学方法，评估金（Au）、氧化铜（CuO）及碲化镉（CdTe）纳米颗粒对先天免疫系统的影响。本研究选用人单核细胞系THP-1作为实验模型，以抗癌药物喜树碱（camptothecin）与阿霉素（doxorubicin）作为细胞死亡的阳性对照，以脂多糖（lipopolysaccharide）作为促炎激活的阳性对照。尽管三种纳米颗粒的整体毒性效应相当（死细胞占比为50 ± 10%），但它们诱导产生的蛋白质组学特征却存在显著差异：碲化镉纳米颗粒的诱导效应最强，其次为氧化铜纳米颗粒与金纳米颗粒。碲化镉纳米颗粒的毒性机制涉及拓扑异构酶（topoisomerases）的下调表达。氧化铜纳米颗粒的效应最类似于氧化应激，其涉及热应激相关蛋白的上调表达。金纳米颗粒可诱导炎症介质核因子κB（NF-κB）的上调表达，其抑制剂TIPE2被确定为金纳米颗粒的直接作用靶点。此外，金纳米颗粒可触发核因子κB的激活，这一点可通过p65亚基的磷酸化得到证实。综上，本研究所述的组合式蛋白质组学方法可用于表征纳米颗粒对免疫细胞的影响。