低剂量锂对神经母细胞瘤细胞系具有神经保护作用

Ş. Güran, Z. Çoban, H. Gündeşli, Özgür Kılıçarslan
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摘要

在神经退行性疾病中,神经元变性是一个重要的过程。本研究的目的是通过考虑几个具有神经保护作用的特定基因来确定一定数量的Li对神经母细胞瘤细胞的影响。制备浓度分别为1 μM、15 μM(低剂量)、30 μM和45 μM(高剂量)的Li溶液,分别作用于神经母细胞瘤细胞系。XTT法和台盼蓝法分别测定细胞增殖和活力。采用定量实时聚合酶链反应(qRT-PCR)检测NES、BDNF、GRIN2A、LRRK2、PRKN、SNCA mRNA表达水平。低剂量Li处理的细胞活力显著增加,而高剂量Li处理的细胞活力与未处理的对照相比显著降低。此外,与对照组相比,高剂量组细胞的增殖率显著降低。结果表明,几个基因(NES、LRRK2、PRKN)的mRNA表达水平显著上调。对于BDNF,仅在极少量Li处理的细胞中,该基因的表达显著上调。然而,没有获得关于GRIN2A的重要数据。此外,与对照组相比,SNCA mRNA表达水平明显下调。在不同浓度的锂离子作用下,细胞内NES、LRRK2、PRKN、BDNF和SNCA基因的表达具有统计学意义,这表明锂离子对神经元存活相关基因的转录调控起作用。这些发现支持剂量依赖性Li治疗可能对神经退行性疾病具有保护作用。
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LITHIUM HAS NEUROPROTECTIVE EFFECT ON NEUROBLASTOMA CELL LINE IN LOW DOSAGES
Lithium (Li) was presented as a protective agent in neuron degeneration which is an important process in neurodegenerative diseases. The aim of this study was to determine the effect of certain amounts of Li on neuroblastoma cells by considering several specific genes which act on neuroprotection. Li solutions were prepared as 1 μM, 15 μM (low dosage), 30 μM and 45 µM (high dosage) concentrations then applied to neuroblastoma cell line. XTT and trypan blue assays were performed to determine the cell proliferation and viability, respectively. mRNA expression levels of NES, BDNF, GRIN2A, LRRK2, PRKN, and SNCA were detected by quantitative real time polymerase chain reaction (qRT-PCR). Cell viability detected as significantly increased in cells treated with low dosage Li however, it was significantly decreased in high dosage applied cells compared to untreated control. In addition, cell proliferation ratios were significantly decreased in high dosage applied cells compared to control. It was demonstrated that mRNA expression levels of several genes (NES, LRRK2, PRKN) were significantly upregulated. Regarding to BDNF, expression of the gene was significantly upregulated in the cells only treated with very low amount of Li. However, no significant data could be obtained for GRIN2A. Furthermore, mRNA expression level of SNCA was determined as significantly downregulated compared to control. Statistically significant expression of NES, LRRK2, PRKN, BDNF and SNCA genes due to the variable Li concentrations applied to cells suggests that Li acts on transcriptional regulation of certain genes associated with neuronal survival. These findings support that dose dependent Li treatment might have a protective effect for neurodegenerative diseases.
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