Ultrafine particles cause cytoskeletal dysfunctions in macrophages
Ultrafine particles cause cytoskeletal dysfunctions in macrophages
复制标题
DOI：
10.1006/taap.2002.9430
复制
发表时间：
2002-08-01
影响因子：
3.8
通讯作者：
Heyder, J
中科院分区：
文献类型：
--
作者：
Möller, W;Hofer, T;Heyder, J
巨噬细胞的基本细胞骨架功能是迁移、吞噬异物以及细胞内运输和消化。细和超细测试颗粒（UFP），如TiO 2、元素碳、商业炭黑、柴油机排气颗粒物和城市灰尘（UrbD），对巨噬细胞的细胞骨架相关功能的影响，如吞噬作用、吞噬体运输机制和机械细胞骨架完整性，通过流式细胞术和细胞磁性测定法进行研究。此外，检测到由测试颗粒引起的坏死和凋亡。测试颗粒的直径范围为12至220 nm，Brunauer-Emmet-Teller比表面积范围为6至600 m2/g。使用通过支气管肺泡灌洗获得的比格犬（BD-AM）的原代肺泡巨噬细胞以及源自细胞系J774A.1的巨噬细胞。对于cytomagnetometry研究，球形1.8 μ m铁磁颗粒作为细胞骨架功能的探针，并与巨噬细胞一起孵育24小时前UFP曝光。将巨噬细胞在体外暴露于10-320 μ g UFP/ml/10（6）细胞长达24小时。在所有实验中，J774A.1巨噬细胞比BD-AM对UFP暴露更敏感。通过细胞磁性测定法评价的细胞骨架功能障碍是吞噬体转运受损和细胞骨架刚度增加，并在100 μ g UFP/ml/106细胞及以上浓度下发生，在BD-AM和J774A.1中。只有细TiO 2没有显示出任何效果。城市灰尘（标准参考物质1649 a）和柴油机废气颗粒（DEP，标准参考物质1650）引起的细胞骨架功能障碍与元素碳具有高比表面积。用二氯甲烷洗涤颗粒后，DEP或UrbD诱导的细胞骨架功能障碍可以减轻。UFP导致1-μ m直径的荧光乳胶珠的吞噬功能受损，抑制细胞增殖，并降低细胞活力。所有记录的细胞毒性参数仅显示与比表面积或UFP总数的弱相关性，这可能是由于不同类型的颗粒和不同的表面组成。UFP在体外引起巨噬细胞的细胞骨架毒性，这可引起细胞功能障碍，如增殖受损、吞噬活性受损、细胞内转运过程延迟以及细胞硬度增加，并可导致肺防御能力受损。(C)2002 Elsevier Science（美国）。
Essential cytoskeletal functions of macrophages are migration, phagocytosis of foreign materials, and intracellular transport and digestion The influence of fine and ultrafine test particles (UFP), such as TiO2, elemental carbon, commercial carbon black, diesel exhaust particulate matter, and urban dust (UrbD), on cytoskeleton-related functions of macrophages, such as phagocytosis, phagosome transport mechanisms, and mechanical cytoskeletal integrity, were studied by flow cytometry and by cytomagnetometry. Additionally, necrosis and apoptosis caused by the test particles was detected. The diameter of the test particles ranged from 12 to 220 nm and the Brunauer-Emmet-Teller specific surface area ranged from 6 to 600 m(2)/g. Primary alveolar macrophages from beagle dogs (BD-AM), obtained by bronchoalveolar lavage, were used as well as macrophages originating from the cell line J774A.1. For cytomagnetometry studies, spherical 1.8-mum ferromagnetic particles served as probes for cytoskeletal functions and were incubated together with the macrophages 24 h prior to UFP exposure. Macrophages were exposed in vitro with 10-320 mug UFP/ml/10(6) cells up to 24 h. In all experiments, J774A.1 macrophages were more sensitive than BD-AM to UFP exposure. Cytoskeletal dysfunctions evaluated by cytomagnetometry were an impaired phagosome transport and an increased cytoskeletal stiffness and occurred at concentrations of 100 mug UFP/ml/106 cells and above, in both BD-AM and J774A.1. Only fine TiO2 did not show any effect. Urban dust (standard reference material 1649a) and diesel exhaust particles (DEP, standard reference material 1650) caused comparable cytoskeletal dysfunctions to elemental carbon with high specific surface area. Cytoskeletal dysfunctions induced by DEP or UrbD could be reduced after washing the particles by dichloromethane. UFP caused an impaired phagocytosis of 1-mum diameter fluorescent latex beads, inhibited cell proliferation, and decreased cell viability. All recorded cytotoxic parameters showed only weak correlations with the specific surface area or the total number of UFP, which can result from the different types of particles and different surface compositions. UFP cause cytoskeletal toxicity in vitro in macrophages, which can cause cellular dysfunctions, such as impaired proliferation, impaired phagocytic activity, and retarded intracellular transport processes as well as increased cell stiffness and can result in impaired defense ability in the lung. (C) 2002 Elsevier Science (USA).