Oberemm Axel, Hansen Ulf, Böhmert Linda, Meckert Christine, Braeuning Albert, Thünemann Andreas F, Lampen Alfonso
Federal Institute for Risk Assessment, Max-Dohrn Str. 8-10, 10589, Berlin, Germany.
Federal Institute for Materials Research and Testing, Unter den Eichen 87, 12205, Berlin, Germany.
J Appl Toxicol. 2016 Mar;36(3):404-13. doi: 10.1002/jat.3231. Epub 2015 Oct 5.
Even although quite a number of studies have been performed so far to demonstrate nanoparticle-specific effects of substances in living systems, clear evidence of these effects is still under debate. The present study was designed as a comparative proteomic analysis of human intestinal cells exposed to a commercial silver nanoparticle reference material and ions from AgNO3. A two-dimensional gel electrophoresis/MALDI mass spectrometry (MS)-based proteomic analysis was conducted after 24-h incubation of differentiated Caco-2 cells with non-cytotoxic and low cytotoxic silver concentrations (2.5 and 25 µg ml(-1) nanosilver, 0.5 and 5 µg ml(-1) AgNO3). Out of an overall number of 316 protein spots differentially expressed at a fold change of ≥ 1.4 or ≤ -1.4 in all treatments, 169 proteins could be identified. In total, 231 spots were specifically deregulated in particle-treated groups compared with 41 spots, which were limited to AgNO3-treatments. Forty-four spots (14 %) were commonly deregulated by both types of treatment. A considerable fraction of the proteins differentially expressed after treatment with nanoparticles is related to protein folding, synthesis or modification of proteins as well as cellular assembly and organization. Overlays of networks obtained for particulate and ionic treatments showed matches, indicating common mechanisms of combined particle and ionic silver exposure and exclusive ionic silver treatment. However, proteomic responses of Caco-2 cells treated with higher concentrations of silver species also showed some differences, for example regarding proteins related to fatty acid and energy metabolism, suggesting an induction of also some different molecular mechanisms for particle exposure and ionic treatment.
尽管到目前为止已经进行了相当多的研究来证明生物系统中物质的纳米颗粒特异性效应,但这些效应的明确证据仍在争论中。本研究旨在对暴露于商业银纳米颗粒参考材料和硝酸银离子的人肠道细胞进行比较蛋白质组学分析。在用无细胞毒性和低细胞毒性的银浓度(2.5和25μg ml⁻¹纳米银,0.5和5μg ml⁻¹硝酸银)对分化的Caco-2细胞进行24小时孵育后,进行了基于二维凝胶电泳/基质辅助激光解吸电离质谱(MS)的蛋白质组学分析。在所有处理中,以≥1.4或≤ -1.4的倍数变化差异表达的316个蛋白质斑点中,有169个蛋白质可以被鉴定出来。与仅限于硝酸银处理的41个斑点相比,颗粒处理组中共有231个斑点被特异性失调。两种处理共有44个斑点(14%)被失调。纳米颗粒处理后差异表达的相当一部分蛋白质与蛋白质折叠、蛋白质合成或修饰以及细胞组装和组织有关。颗粒处理和离子处理获得的网络叠加显示有匹配,表明颗粒银和离子银联合暴露以及单独离子银处理的共同机制。然而,用较高浓度银物种处理的Caco-2细胞的蛋白质组学反应也显示出一些差异,例如与脂肪酸和能量代谢相关的蛋白质,这表明颗粒暴露和离子处理也诱导了一些不同的分子机制。