Kim Dong-Yung, Kim Jun-Hyung, Lee Jae-Chul, Won Moo-Ho, Yang Se-Ran, Kim Hyoung-Chun, Wie Myung-Bok
Department of Veterinary Toxicology, College of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
Department of Neurobiology, School of Medicine, Kangwon National University, Chuncheon, Korea.
Toxicol Res. 2019 Jan;35(1):83-91. doi: 10.5487/TR.2019.35.1.083. Epub 2018 Jan 15.
Nanoparticles (NPs) have been recognized as both useful tools and potentially toxic materials in various industrial and medicinal fields. Previously, we found that zinc oxide (ZnO) NPs that are neurotoxic to human dopaminergic neuroblastoma SH-SY5Y cells are mediated by lipoxygenase (LOX), not cyclooxygenase-2 (COX-2). Here, we examined whether human bone marrow-derived mesenchymal stem cells (MSCs), which are different from neuroblastoma cells, might exhibit COX-2- and/or LOX-dependent cytotoxicity of ZnO NPs. Additionally, changes in annexin V expression, caspase-3/7 activity, and mitochondrial membrane potential (MMP) induced by ZnO NPs and ZnO were compared at 12 hr and 24 hr after exposure using flow cytometry. Cytotoxicity was measured based on lactate dehydrogenase activity and confirmed by trypan blue staining. Rescue studies were executed using zinc or iron chelators. ZnO NPs and ZnO showed similar dose-dependent and significant cytotoxic effects at concentrations ≥ 15 μg/mL, in accordance with annexin V expression, caspase-3/7 activity, and MMP results. Human MSCs exhibited both COX-2 and LOX-mediated cytotoxicity after exposure to ZnO NPs, which was different from human neuroblastoma cells. Zinc and iron chelators significantly attenuated ZnO NPs-induced toxicity. Conclusively, these results suggest that ZnO NPs exhibit both COX-2- and LOX-mediated apoptosis by the participation of mitochondrial dysfunction in human MSC cultures.
纳米颗粒（NPs）在各种工业和医学领域中既被视为有用的工具，也被视为潜在的有毒物质。此前，我们发现对人多巴胺能神经母细胞瘤SH-SY5Y细胞具有神经毒性的氧化锌（ZnO）纳米颗粒是由脂氧合酶（LOX）介导的，而非环氧合酶-2（COX-2）。在此，我们研究了与神经母细胞瘤细胞不同的人骨髓间充质干细胞（MSCs）是否可能表现出ZnO纳米颗粒的COX-2和/或LOX依赖性细胞毒性。此外，使用流式细胞术比较了暴露后12小时和24小时时ZnO纳米颗粒和ZnO诱导的膜联蛋白V表达、半胱天冬酶-3/7活性和线粒体膜电位（MMP）的变化。基于乳酸脱氢酶活性测量细胞毒性，并通过台盼蓝染色进行确认。使用锌或铁螯合剂进行挽救研究。根据膜联蛋白V表达、半胱天冬酶-3/7活性和MMP结果，ZnO纳米颗粒和ZnO在浓度≥15μg/mL时表现出相似的剂量依赖性和显著的细胞毒性作用。人骨髓间充质干细胞在暴露于ZnO纳米颗粒后表现出COX-2和LOX介导的细胞毒性，这与人神经母细胞瘤细胞不同。锌和铁螯合剂显著减轻了ZnO纳米颗粒诱导的毒性。总之，这些结果表明ZnO纳米颗粒通过线粒体功能障碍参与在人骨髓间充质干细胞培养物中表现出COX-2和LOX介导的细胞凋亡。
