Rodríguez-Yáñez Yury, Bahena-Uribe Daniel, Chávez-Munguía Bibiana, López-Marure Rebeca, González-Monroy Stuart, Cisneros Bulmaro, Albores Arnulfo
Departamento de Toxicología, Centro de Investigación y de Estudios Avanzados del IPN (Cinvestav), Mexico.
Laboratorio Avanzado de Nanoscopía Electrónica (LANE), Cinvestav, Mexico.
Toxicol In Vitro. 2015 Aug;29(5):1201-14. doi: 10.1016/j.tiv.2015.02.009. Epub 2015 Mar 16.
Recent studies have demonstrated that carbon nanotubes (CNTs) induce platelet aggregation, endothelial dysfunction and vascular thrombosis. However, there is little information on the effects of CNTs on fibrinolysis. We investigated the role of pristine-commercial single-walled carbon nanotubes (SWCNTs) with <3% Co content in fibrinolysis and their contribution to the induction of pro-thrombotic processes in human vein endothelial cells (HUVEC). SWCNTs alone produced concentration-dependent oxidation, as measured by a dithiothreitol oxidation assay. Internalized SWCNTs were located in HUVEC treated with 25 μg/ml using transmission electron microscopy, whereas treatment with 50 μg/ml compromised cell viability, and oxidative stress increased significantly at 5 μg/ml. The study showed that in HUVEC treated with 25 μg SWCNT/ml, fibrinolysis-related gene expression and protein levels had increased by 3-12 h after treatment (serpine-1: 13-fold; PLAT: 11-fold and PLAU: 2-fold), but only the PAI-1 protein was increased (1.5-fold), whereas tissue and urokinase plasminogen activator proteins (tPA and uPA, respectively) tended to decrease. In summary, pristine SWCNTs treatment resulted in evident HUVEC damage caused by cell fiber contact, internalization, and oxidative stress due to contaminant metals. The generation of endothelial dysfunction, as shown by the altered expression of genes and proteins involved in fibrinolysis, suggest that SWCNTs display pro-thrombotic effects.
最近的研究表明，碳纳米管（CNTs）可诱导血小板聚集、内皮功能障碍和血管血栓形成。然而，关于碳纳米管对纤维蛋白溶解作用的信息却很少。我们研究了钴含量<3%的原始商业单壁碳纳米管（SWCNTs）在纤维蛋白溶解中的作用及其对人静脉内皮细胞（HUVEC）促血栓形成过程诱导的贡献。通过二硫苏糖醇氧化试验测定，单独的SWCNTs会产生浓度依赖性氧化。使用透射电子显微镜观察发现，内化的SWCNTs存在于用25μg/ml处理的HUVEC中，而用50μg/ml处理会损害细胞活力，并且在5μg/ml时氧化应激显著增加。研究表明，在用25μg SWCNT/ml处理的HUVEC中，处理后3 - 12小时纤维蛋白溶解相关基因表达和蛋白质水平增加（丝氨酸蛋白酶抑制剂-1：13倍；组织型纤溶酶原激活剂：11倍；尿激酶型纤溶酶原激活剂：2倍），但仅纤溶酶原激活物抑制剂-1蛋白增加（1.5倍），而组织型和尿激酶型纤溶酶原激活剂蛋白（分别为tPA和uPA）则趋于减少。总之，原始SWCNTs处理导致明显的HUVEC损伤，这是由细胞纤维接触、内化以及污染物金属引起的氧化应激所致。纤维蛋白溶解相关基因和蛋白质表达的改变表明内皮功能障碍的产生，这提示SWCNTs具有促血栓形成作用。
