Ortega V A, Katzenback B A, Stafford J L, Belosevic M, Goss G G
Department of Biological Sciences .
Nanotoxicology. 2015 Feb;9(1):23-33. doi: 10.3109/17435390.2013.861943. Epub 2013 Dec 9.
Exposure effects from polyacrylic acid (PAA) metal-oxide nanoparticles (TiO2, CeO2, Fe2O3, ZnO) on fish neutrophil viability and effector functions (degranulation, respiratory burst, inflammatory gene expression) were investigated using primary kidney goldfish (Carassius auratus L.) neutrophils as a model. Several studies have reported cytotoxic effects of NPs but there are limited reports on their potential to perturb the innate immune system of aquatic organisms. PAA-TiO2 significantly decreased neutrophil viability in a time and dose-dependent manner at all measured time points (0-48 h) and concentrations (0-200 µg/mL). Maximum viability decreased by (mean ± SEM): 67.1 ± 3.3%, 78.4 ± 4.2% and 74.9 ± 5.0% when exposed to 50, 100 and 200 µg/mL for 48 h, respectively. PAA-ZnO also significantly decreased neutrophil viability but only at 48 h exposures at higher concentrations. Neutrophil degranulation increased by approximately 3% after 30 min and by 8% after 4 h when exposed to sublethal doses (10 µg/mL) of PAA-CeO2 or PAA-Fe2O3. All PAA-NPs induced an increase in neutrophil respiratory burst when exposed to 10 µg/mL for 30 and 60 min, however, PAA-Fe2O3 was the only NP where the response was significant. Lastly, NPs altered the expression of a number of pro-inflammatory and immune genes, where PAA-TiO2 most significantly increased the mRNA levels of pro-inflammatory genes (il-1b, ifng) in neutrophils by 3 and 2.5 times, respectively. Together, these data demonstrate that goldfish neutrophils can be negatively affected from exposures to PAA-coated NPs and are functionally responsive to specific core-material properties at sublethal doses. These changes could perturb the innate response and affect the ability of fish to respond to pathogens.
以原代金鱼（Carassius auratus L.）中性粒细胞为模型，研究了聚丙烯酸（PAA）金属氧化物纳米颗粒（TiO2、CeO2、Fe2O3、ZnO）对鱼类中性粒细胞活力和效应功能（脱颗粒、呼吸爆发、炎症基因表达）的暴露影响。多项研究报道了纳米颗粒的细胞毒性作用，但关于它们扰乱水生生物先天免疫系统潜力的报道有限。PAA-TiO2在所有测量时间点（0 - 48小时）和浓度（0 - 200μg/mL）下，均以时间和剂量依赖的方式显著降低中性粒细胞活力。当分别暴露于50、100和200μg/mL 48小时时，最大活力分别降低了（平均值±标准误）：67.1±3.3%、78.4±4.2%和74.9±5.0%。PAA-ZnO也显著降低中性粒细胞活力，但仅在较高浓度下暴露48小时时出现。当暴露于亚致死剂量（10μg/mL）的PAA-CeO2或PAA-Fe2O3时，中性粒细胞脱颗粒在30分钟后增加约3%，4小时后增加8%。所有PAA纳米颗粒在暴露于10μg/mL 30和60分钟时均诱导中性粒细胞呼吸爆发增加，然而，PAA-Fe2O3是唯一反应显著的纳米颗粒。最后，纳米颗粒改变了许多促炎和免疫基因的表达，其中PAA-TiO2最显著地使中性粒细胞中促炎基因（il-1b、ifng）的mRNA水平分别增加了３倍和2.5倍。总之，这些数据表明，金鱼中性粒细胞可能会受到暴露于PAA包被纳米颗粒的负面影响，并且在亚致死剂量下对特定核心材料特性具有功能反应。这些变化可能会扰乱先天反应并影响鱼类对病原体的反应能力。
