Department of Medical Laboratory, Özalp Vocational High SchoolVan Yuzuncu Yil University, Van, Turkey.
Department of Biochemistry, Faculty of Veterinary Medicine, Van Yuzuncu Yil University, Van, Turkey.
Mol Biol Rep. 2020 Oct;47(10):7761-7770. doi: 10.1007/s11033-020-05852-2. Epub 2020 Oct 6.
Prolonged and excessive fluoride exposure can lead to fluorosis. The kidney is one of the organs that are injured mostly due to fluoride-induced damage. Fluoride can induce DNA damage at cytotoxic concentrations. This study aims to determine the extent of NaF-induced DNA damage and to investigate the effect of vitamin E and selenium combination (ES) in preventing and repairing this damage. For this purpose, we administered different combinations of NaF and ES to NRK-52E cells and determined the effective concentrations of ES and the NaF IC values associated with different incubation times (3, 12, and 24 h) by using the MTT assay. The determined quantities of NaF IC in association with time and the NaF IC + ES combination were administered to the cells. The extent of DNA damage was determined with the comet assay and the expression levels of the Ku70/80 and PARP-1 genes were determined with the RT-qPCR method. DNA damage significantly increased in all experimental groups compared to the control group (p < 0.05). It was found out that the NaF and ES combination statistically reduced the DNA damage compared to the damage observed in the NaF-treated groups (p < 0.05). Treatment of the ES combination significantly increased the expressions of Ku70 and Ku80 genes involved in DNA repair (p < 0.05). We concluded that vitamin E and selenium can potentially be effective in the repair of fluoride-induced DNA damage based on the results of this in vitro study. Our results may shed light on the prevention of DNA damage associated with fluorosis.
长期和过量的氟暴露会导致氟中毒。肾脏是受氟化物损伤影响最大的器官之一。氟化物在细胞毒性浓度下会引起 DNA 损伤。本研究旨在确定 NaF 诱导的 DNA 损伤的程度,并研究维生素 E 和硒联合(ES)在预防和修复这种损伤中的作用。为此,我们用不同组合的 NaF 和 ES 处理 NRK-52E 细胞,并通过 MTT 测定法确定 ES 的有效浓度和与不同孵育时间(3、12 和 24 h)相关的 NaF IC 值。用时间和 NaF IC+ES 组合确定的 NaF IC 定量值被施用于细胞。彗星试验测定 DNA 损伤程度,RT-qPCR 法测定 Ku70/80 和 PARP-1 基因的表达水平。与对照组相比,所有实验组的 DNA 损伤均显著增加(p<0.05)。结果表明,与 NaF 处理组观察到的损伤相比,NaF 和 ES 联合显著降低了 DNA 损伤(p<0.05)。ES 联合治疗显著增加了参与 DNA 修复的 Ku70 和 Ku80 基因的表达(p<0.05)。基于这项体外研究的结果,我们得出结论,维生素 E 和硒可能对修复氟化物诱导的 DNA 损伤有效。我们的结果可能为预防氟中毒相关的 DNA 损伤提供线索。