甲醇致SK-N-SH细胞氧化应激及DNA损伤的研究

Oxidative Stress and DNA Damage Induced by Methanol in SK-N-SH Cells

目的通过甲醇对SK-N-SH细胞染毒,探讨SK-N-SH细胞氧化应激与DNA损伤情况及二者的关系。方法选取体外培养的SK-N-SH细胞,通过CCK-8法确定染毒剂量和染毒时间。采用不同浓度甲醇(0、250、750、1250 mmol·L^-1)分别染毒细胞24 h后,用活性氧(ROS)试剂盒检测ROS水平;单细胞琼脂糖凝胶电泳检测DNA损伤情况。结果甲醇染毒24 h后,ROS水平中、高浓度组高于对照组,高浓度组高于低浓度组(P均<0.05)。DNA平均拖尾长度及拖尾率低、中、高浓度组高于对照组,中、高浓度组高于低浓度组(P均<0.01)。ROS水平与DNA平均拖尾长度呈正相关关系(r=0.818,P<0.01)。结论甲醇能诱导SK-NSH细胞发生氧化应激与DNA损伤反应,其可能的机制是通过诱导SK-N-SH细胞发生氧化应激后产生DNA损伤。

Objective Through methanol poisoning of SK-N-SH cells,the relationship of oxidative stress and DNA damage of methanol-induced SK-N-SH cells was studied.Methods In vitro cultured SK-N-SH cells were selected,the dose and time of exposure were determined by CCK-8 method.After treatment with 0,250,750,and 1250 mmol·L^-1 of methanol(control,low,medium and high methanol concentration groups)for 24 hours,ROS level was detected by reactive oxygen species(ROS)detection kit,and DNA damage was detected by single cell agarose gelelectrophoresis.Results After 24 h,the ROS levels in the medium and high concentration groups were higher than that of the control group(P all<0.05),and the high concentration group was higher than the low concentration group(P<0.05).The average tailing length and tailing rate of the low,medium and high concentration groups were significantly higher than the control group(P all<0.01).The medium and high concentration groups were significantly higher than the low concentration group(P all<0.01).There was a positive correlation between ROS level and the average tail length of DNA(r=0.818,P<0.01).Conclusion Methanol can induce oxidative stress and DNA damage in SK-N-SH cells.The possible mechanism is that DNA damage is induced by oxidative stress in SK-N-SH cells.