不同直径的非功能化聚苯乙烯纳米颗粒对人外周血单个核细胞凋亡和 mTOR 水平的诱导作用。

The effects of non-functionalized polystyrene nanoparticles of different diameters on the induction of apoptosis and mTOR level in human peripheral blood mononuclear cells.

机构信息

University of Lodz, Faculty of Biology and Environmental Protection, Department of Biophysics of Environmental Pollution, 141/143 Pomorska St., 90-236, Lodz, Poland.

出版信息

Chemosphere. 2023 Sep;335:139137. doi: 10.1016/j.chemosphere.2023.139137. Epub 2023 Jun 5.

DOI:10.1016/j.chemosphere.2023.139137

PMID:37285979

Abstract

Particles of various types of plastics, including polystyrene nanoparticles (PS-NPs), have been determined in human blood, placenta, and lungs. These findings suggest a potential detrimental effect of PS-NPs on bloodstream cells. The purpose of this study was to assess the mechanism underlying PS-NPs-induced apoptosis in human peripheral blood mononuclear cells (PBMCs). Non-functionalized PS-NPs of three diameters: 29 nm, 44 nm, and 72 nm were studied used in this research. PBMCs were isolated from human leukocyte-platelet buffy coat and treated with PS-NPs at concentrations ranging from 0.001 to 200 μg/mL for 24 h. Apoptotic mechanism of action was evaluated by determining the level of cytosolic calcium ions, as well as mitochondrial transmembrane potential, and ATP levels. Furthermore, detection of caspase-8, -9, and -3 activation, as well as mTOR level was conducted. The presence of apoptotic PBMCs was confirmed by the method of double staining of the cells with propidium iodide and FITC-conjugated Annexin V. We found that all tested NPs increased calcium ion and depleted mitochondrial transmembrane potential levels. The tested NPs also activated caspase-9 and caspase-3, and the smallest NPs of 29 nm of diameter also activated caspase-8. The results clearly showed that apoptotic changes and an increase of mTOR level depended on the size of the tested NPs, while the smallest particles caused the greatest alterations. PS-NPs of 26 nm of diameter activated the extrinsic pathway (increased caspase-8 activity), as well as intrinsic (mitochondrial) pathway (increased caspase-9 activity, raised calcium ion level, and decreased transmembrane mitochondrial potential) of apoptosis. All PS-NPs increased mTOR level at the concentrations smaller than those that induced apoptosis and its level returned to control value when the process of apoptosis escalated.

摘要

各种类型的塑料颗粒，包括聚苯乙烯纳米颗粒（PS-NPs），已在人体血液、胎盘和肺部中被发现。这些发现表明 PS-NPs 对血液细胞可能有潜在的有害影响。本研究旨在评估 PS-NPs 诱导人外周血单个核细胞（PBMCs）凋亡的机制。本研究使用三种直径的非功能化 PS-NPs：29nm、44nm 和 72nm。从人白细胞-血小板缓冲层中分离 PBMCs，并在浓度范围为 0.001 至 200μg/mL 的 PS-NPs 下处理 24 小时。通过测定细胞质钙离子水平、线粒体跨膜电位和 ATP 水平来评估凋亡作用机制。此外，还检测了 caspase-8、-9 和 -3 的激活以及 mTOR 水平。通过碘化丙啶和 FITC 缀合的 Annexin V 双重染色细胞的方法来确认凋亡 PBMCs 的存在。我们发现所有测试的 NPs 均增加了钙离子并耗竭了线粒体跨膜电位。测试的 NPs 还激活了 caspase-9 和 caspase-3，而直径 29nm 的最小 NPs 还激活了 caspase-8。结果清楚地表明，凋亡变化和 mTOR 水平的增加取决于所测试的 NPs 的大小，而最小的颗粒会引起最大的变化。直径 26nm 的 PS-NPs 激活了外源性途径（增加 caspase-8 活性）以及内源性（线粒体）途径（增加 caspase-9 活性、提高钙离子水平和降低跨膜线粒体电位）的凋亡。所有 PS-NPs 在引起凋亡的浓度以下均增加了 mTOR 水平，当凋亡过程加剧时，其水平恢复到对照值。