Center for Medical Research, Medical University of Graz, Austria.
Part Fibre Toxicol. 2012 Jul 12;9:26. doi: 10.1186/1743-8977-9-26.
Data from environmental exposure to nanoparticles (NPs) suggest that chronic exposure may increase the incidence of lung, cardiovascular and neurodegenerative diseases. Impairment of cell function by intracellular accumulation of NPs is also suspected. Many types of NPs have been detected in the endosomal-lysosomal system and, upon repeated exposure, alterations of the endosomal-lysosomal system may occur. To identify such effects we compared the effect of carboxyl polystyrene particles (CPS) of different sizes (20-500 nm) on lysosomes of the endothelial cell line EAhy926 after short (24h) and long (72h-96h) exposure times. Lysosomal localization of CPS, as well as lysosomal pH, lysosomal membrane integrity, morphology of the endosomal-lysosomal system and activities of the lysosomal enzymes,cathepsin B and sulfatases, upon exposure to CPS were recorded.
CPS in sizes ≤100 nm showed high co-localization with lysosomes already after 4h, larger CPS after 24h. None of the particles at non-cytotoxic concentrations caused marked changes in lysosomal pH or destroyed lysosomal membrane integrity. At 24h of exposure, 20 nm CPS induced significant dilatation of the endosomal-lysosomal system and reduced activity of lysosomal sulfatases. After 72h, these alterations were less pronounced.
Despite accumulation in lysosomes CPS induced only small changes in lysosomes. Upon longer contact, these changes are even less pronounced. The presented panel of assays may serve to identify effects on lysosomes also for other NPs.
来自于纳米颗粒(NPs)环境暴露的数据表明,慢性暴露可能会增加肺部、心血管和神经退行性疾病的发病率。细胞内 NP 积累导致细胞功能受损也受到怀疑。多种类型的 NPs 已在内体溶酶体系统中被检测到,并且在反复暴露后,内体溶酶体系统可能会发生改变。为了确定这些影响,我们比较了不同大小(20-500nm)的羧基聚苯乙烯颗粒(CPS)对内皮细胞系 EAhy926 的溶酶体的短期(24h)和长期(72h-96h)暴露后的影响。记录了 CPS 对内体溶酶体系统的溶酶体定位、溶酶体 pH、溶酶体膜完整性、内体溶酶体系统的形态以及溶酶体酶、组织蛋白酶 B 和硫酸酯酶的活性的影响。
尺寸≤100nm 的 CPS 在 4h 后就与溶酶体高度共定位,较大的 CPS 在 24h 后才与溶酶体共定位。在非细胞毒性浓度下,没有一种颗粒会显著改变溶酶体 pH 或破坏溶酶体膜完整性。在暴露 24h 时,20nm 的 CPS 会导致内体溶酶体系统明显扩张,并降低溶酶体硫酸酯酶的活性。72h 后,这些变化不那么明显。
尽管 CPS 在溶酶体中积累,但仅对溶酶体产生较小的影响。在更长的接触时间内,这些变化甚至更小。本研究提出的检测方法可以用于识别其他 NPs 对内体溶酶体的影响。