a Department of Toxicology and Sanitary Chemistry, School of Public Health , Capital Medical University , Beijing , P.R. China.
b Beijing Key Laboratory of Environmental Toxicology , Capital Medical University , Beijing , P.R. China.
Nanotoxicology. 2018 Jun;12(5):470-484. doi: 10.1080/17435390.2018.1461267. Epub 2018 Apr 14.
Nowadays, nanotechnology environmental health and safety (nanoEHS) is gaining attention. We previously found that silica nanoparticles (SiNPs) could induce vascular endothelial damage. However, the subsequent toxicologic response to SiNPs-induced endothelial damage was still largely unknown. In this study, we explored the inflammation-coagulation response and thrombotic effects of SiNPs in endothelial cells and zebrafish embryos. For in vitro study, swollen mitochondria and autophagosome were observed in ultrastructural analysis. The cytoskeleton organization was disrupted by SiNPs in vascular endothelial cells. The release of proinflammatory and procoagulant cytokines including IL-6, IL-8, MCP-1, PECAM-1, TF and vWF, were markedly elevated in a dose-dependent manner. For in vivo study, based on the NOAEL for dosimetry selection, and using two transgenic zebrafish, Tg(mpo:GFP) and Tg(fli-1:EGFP), SiNPs-induced neutrophil-mediated inflammation and impaired vascular endothelial cells. With the dosage higher than NOAEL, SiNPs significantly decreased blood flow and velocity, exhibiting a blood hypercoagulable state in zebrafish embryos. The thrombotic effect was assessed by o-dianisidine staining, showed that an increasing of erythrocyte aggregation occurred in SiNPs-treated zebrafish. Microarray analysis was used to screen the possible genes for inflammation-coagulation response to SiNPs in zebrafish, and the JAK1/TF signaling pathway was further verified by qRT-PCR and Western blot assays. For in-deepth study, il6st was knocked down with specific morpholinos. The whole-mount in situ hybridization and qRT-PCR analysis showed that the expression jak1 and f3b were attenuated in il6st knockdown groups. In summary, our data demonstrated that SiNPs could induce inflammation-coagulation response and thrombotic effects via JAK1/TF signaling pathway.
如今,纳米技术的环境健康与安全(nanoEHS)正受到关注。我们之前发现,二氧化硅纳米颗粒(SiNPs)可诱导血管内皮损伤。然而,SiNPs 诱导的内皮损伤后的后续毒理学反应在很大程度上仍不清楚。在这项研究中,我们探索了 SiNPs 在血管内皮细胞和斑马鱼胚胎中的炎症-凝血反应和血栓形成效应。在体外研究中,超微结构分析观察到肿胀的线粒体和自噬体。SiNPs 破坏了血管内皮细胞的细胞骨架组织。SiNPs 以剂量依赖性方式显著上调促炎和促凝细胞因子的释放,包括白细胞介素 6(IL-6)、白细胞介素 8(IL-8)、单核细胞趋化蛋白 1(MCP-1)、血小板内皮细胞黏附分子 1(PECAM-1)、组织因子(TF)和血管性血友病因子(vWF)。在体内研究中,基于剂量选择的无可见不良反应水平(NOAEL),并使用两种转基因斑马鱼,髓过氧化物酶:GFP(Tg(mpo:GFP))和 fli-1:EGFP(Tg(fli-1:EGFP)),SiNPs 诱导中性粒细胞介导的炎症和血管内皮细胞损伤。当剂量高于 NOAEL 时,SiNPs 显著降低了斑马鱼胚胎的血流速度,表现出血液高凝状态。通过邻联茴香胺染色评估血栓形成效应,结果表明 SiNPs 处理的斑马鱼中红细胞聚集增加。使用微阵列分析筛选 SiNPs 诱导斑马鱼炎症-凝血反应的可能基因,并用 qRT-PCR 和 Western blot 分析进一步验证 JAK1/TF 信号通路。在深入研究中,用特异性的 morpholino 敲低 il6st。全胚胎原位杂交和 qRT-PCR 分析显示,在 il6st 敲低组中 jak1 和 f3b 的表达减弱。总之,我们的数据表明,SiNPs 可通过 JAK1/TF 信号通路诱导炎症-凝血反应和血栓形成效应。