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二氧化锰纳米颗粒在人神经细胞中诱导的氧化应激所致DNA损伤

Oxidative Stress-Induced DNA Damage by Manganese Dioxide Nanoparticles in Human Neuronal Cells.

作者信息

Alarifi Saud, Ali Daoud, Alkahtani Saad

机构信息

Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.

出版信息

Biomed Res Int. 2017;2017:5478790. doi: 10.1155/2017/5478790. Epub 2017 May 17.

Abstract

Metal nanoparticles have been extensively used in industry as well as in biomedical application. In this work, we have evaluated the toxic potential of manganese dioxide (MnO) nanoparticles (MNPs) on human neuronal (SH-SY5Y) cells. Cellular toxicity due to MNPs (0, 10, 30, and 60 g/ml) on the SH-SY5Y cell was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) tests. MNPs produced reactive oxygen species (ROS) and declined in mitochondrial membrane potential in the SH-SY5Y cell in dose and duration dependent manner. Moreover, lipid peroxide (LPO), superoxide dismutase (SOD), and catalase (CAT) activities were increased and glutathione was reduced in dose and time dependent manner. A significant upgrade in Hoechst 33342 fluorescence intensity (chromosome condensation) and phosphatidylserine translocation (apoptotic cell) was visualized in cells treated with MNPs for 48 h. On the other hand, caspase-3 activity was increased due to MNPs in SH-SY5Y cells. DNA strand breaks were determined by alkaline single cell gel electrophoresis assay (Comet Assay) and maximum fragmentation of DNA produced due to MNPs (60 g/ml) for 48 hours. This result provides a basic mechanism of induction of apoptosis and toxicity by MNPs in SH-SY5Y cells.

摘要

金属纳米颗粒已在工业以及生物医学应用中得到广泛使用。在这项工作中,我们评估了二氧化锰(MnO)纳米颗粒(MNPs)对人神经元(SH-SY5Y)细胞的潜在毒性。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)和中性红摄取(NRU)试验观察了MNPs(0、10、30和60μg/ml)对SH-SY5Y细胞的细胞毒性。MNPs以剂量和持续时间依赖性方式在SH-SY5Y细胞中产生活性氧(ROS)并导致线粒体膜电位下降。此外,脂质过氧化物(LPO)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性以剂量和时间依赖性方式增加,而谷胱甘肽减少。在用MNPs处理48小时的细胞中,观察到Hoechst 33342荧光强度(染色体凝聚)和磷脂酰丝氨酸易位(凋亡细胞)显著升高。另一方面,MNPs导致SH-SY5Y细胞中caspase-3活性增加。通过碱性单细胞凝胶电泳试验(彗星试验)测定DNA链断裂情况,发现MNPs(60μg/ml)处理48小时导致DNA产生最大程度的片段化。该结果提供了MNPs在SH-SY5Y细胞中诱导凋亡和毒性的基本机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/20ad/5449756/1cb5e77e9f99/BMRI2017-5478790.001.jpg

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