Manganese-induced oxidative DNA damage in neuronal SH-SY5Y cells: attenuation of thymine base lesions by glutathione and N-acetylcysteine.
Manganese-induced oxidative DNA damage in neuronal SH-SY5Y cells: attenuation of thymine base lesions by glutathione and N-acetylcysteine.
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DOI：
10.1016/j.toxlet.2012.12.024
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发表时间：
2013-04-26
影响因子：
3.5
通讯作者：
Renee Reams R
中科院分区：
文献类型：
--
作者：
Stephenson AP;Schneider JA;Nelson BC;Atha DH;Jain A;Soliman KF;Aschner M;Mazzio E;Renee Reams R
锰（Mn）是人体正常功能和发育所必需的微量元素。然而，接触高浓度锰可能导致锰中毒，锰中毒是一种进行性神经退行性疾病，其神经症状与特发性帕金森病（IPD）相似。由于暴露于父母的营养、矿山和冶炼厂的蒸气以及焊接烟雾中，锰的身体负担增加与神经健康问题有关。锰神经毒性的潜在机制尚不清楚。因此，本研究旨在研究Mn2+对人神经母细胞瘤SH-SY5Y细胞的毒性作用。与对照相比，Mn2+引起SH-SY5Y细胞活力的浓度依赖性降低。LD50值为12.98 μM Mn2+（对照组与24h Mn处理相比p <0.001）。TUNEL和膜联蛋白V/碘化丙啶细胞凋亡实验证实了Mn2+ （2 μM、62 μM和125 μM）对细胞凋亡的诱导作用。此外，Mn2+诱导DNA单链断裂的形成和积累（通过碱性彗星分析）和氧化修饰胸腺嘧啶碱基（通过气相色谱/质谱分析）。用1mm n -乙酰半胱氨酸或1mm谷胱甘肽等抗氧化剂对细胞进行预孵育，可降低DNA链断裂水平和胸腺嘧啶碱基损伤的形成，提示对氧化性细胞损伤具有保护作用。我们的研究结果表明：1)SH-SY5Y细胞暴露于Mn环境中促进了氧化DNA核苷酸碱基损伤的形成和积累；2)积累DNA损伤的SH-SY5Y细胞更有可能通过凋亡途径死亡；3)添加外源性化学抗氧化剂可以消除积累的DNA损伤水平。这是已知的第一个关于Mn2+诱导SH-SY5Y细胞系胸腺嘧啶病变和抗氧化保护的报道，并为抗氧化剂作为一种治疗策略来保护Mn2+诱导的氧化DNA损伤的潜在应用提供了新的信息。
Manganese (Mn) is an essential trace element required for normal function and development. However, exposure to this metal at elevated levels may cause manganism, a progressive neurodegenerative disorder with neurological symptoms similar to idiopathic Parkinson’s disease (IPD). Elevated body burdens of Mn from exposure to parental nutrition, vapors in mines and smelters and welding fumes have been associated with neurological health concerns. The underlying mechanism of Mn neurotoxicity remains unclear. Accordingly, the present study was designed to investigate the toxic effects of Mn2+ in human neuroblastoma SH-SY5Y cells. Mn2+ caused a concentration dependent decrease in SH-SY5Y cellular viability compared to controls. The LD50 value was 12.98 μM Mn2+ (p <0.001 for control vs. 24h Mn treatment). Both TUNEL and annexin V/propidium iodide apoptosis assays confirmed the induction of apoptosis in the cells following exposure to Mn2+ (2 μM, 62 μM or 125 μM). In addition, Mn2+ induced both the formation and accumulation of DNA single strand breaks (via alkaline comet assay analysis) and oxidatively modified thymine bases (via gas chromatography/mass spectrometry analysis). Pre-incubation of the cells with characteristic antioxidants, either 1 mM N-acetylcysteine or 1 mM glutathione reduced the level of DNA strand breaks and the formation of thymine base lesions, suggesting protection against oxidative cellular damage. Our findings indicate that 1) exposure of SH-SY5Y cells to Mn promotes both the formation and accumulation of oxidative DNA nucleotide base damage, 2) SH-SY5Y cells with accumulated DNA damage are more likely to die via an apoptotic pathway and 3) the accumulated levels of DNA damage can be abrogated by the addition of exogenous chemical antioxidants. This is the first known report of Mn2+-induction and antioxidant protection of thymine lesions in this SH-SY5Y cell line and contributes new information to the potential use of antioxidants as a therapeutic strategy for protection against Mn2+-induced oxidative DNA damage.
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影响因子：
4.4
作者：
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